{"title":"Cell Biology Kit","description":null,"products":[{"product_id":"caspase-1-microplate-assay-kit","title":"Caspase-1 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-1 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299318859,"sku":"abs580189-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/1c481d3d6282401b9206af58c92447c2.jpg?v=1789732979"},{"product_id":"caspase-2-microplate-assay-kit","title":"Caspase-2 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-2 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299384395,"sku":"abs580190-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/eb1de6122a57402888fa2c4f42fd51e7.jpg?v=1789732976"},{"product_id":"caspase-3-microplate-assay-kit","title":"Caspase-3 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-3 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299417163,"sku":"abs580191-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/1e634df20d7e458e83be4f6ba1385de3.jpg?v=1789732980"},{"product_id":"caspase-4-microplate-assay-kit","title":"Caspase-4 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-4 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299482699,"sku":"abs580192-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/9a5b482e9ecd40eabb13aac7d8d84ee0.jpg?v=1789732980"},{"product_id":"caspase-5-microplate-assay-kit","title":"Caspase-5 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-5 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299548235,"sku":"abs580193-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/5606218b796546b6babac47d200abfe9.jpg?v=1789732983"},{"product_id":"caspase-6-microplate-assay-kit","title":"Caspase-6 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-6 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299581003,"sku":"abs580194-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/45bbf5fb8bc64b3b8c0481315a3bd127.jpg?v=1789732984"},{"product_id":"caspase-8-microplate-assay-kit","title":"Caspase-8 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-8 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299679307,"sku":"abs580195-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/1869a4e8385343c2917de8dff7284d0b.jpg?v=1789732989"},{"product_id":"caspase-9-microplate-assay-kit","title":"Caspase-9 Microplate Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eColorimetric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe Dye in the kit is easy to oxidise and deteriorate after dissolving, so it needs to be mixed and used on the spot, and is not recommended for storage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTissue extracts, cell lysates, other biological fluid samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransportation and storage at 4 ℃, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0.01 umol\/ml - 5 umol\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eUsed for the detection and quantification of Caspase-9 activity.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41293299712075,"sku":"abs580196-96T","price":327.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/1b74c7da648a4027bcc2a9d21315dd1f.jpg?v=1789732988"},{"product_id":"trypan-blue-staining-cell-viability-assay-kit","title":"Trypan Blue Staining Cell Viability Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eFYI:\u003cbr\u003e1. Collect cells:\u003cbr\u003eFor adherent cells, the lower cells were first digested with trypsin and\/or EDTA. For suspended cells, the cells can be collected directly. The collected cells were centrifuged at 1000-2000 g for 1 minute, the supernatant was discarded, and the cells were resuspended with 1 mL or appropriate cell resuspension depending on the amount of cells.  \u003cbr\u003e2. Trypan blue staining:\u003cbr\u003eDraw 100 microliters of resuspended cells into a plastic centrifuge tube, add 100 microliters of trypan blue staining solution (2X), gently mix and stain for 3 minutes (3 minutes of staining is enough, and the staining time can be longer, but it should not exceed 10 minutes).\u003cbr\u003e\u003cstrong\u003eNote: For cells resuspended in cell culture medium, an equal volume of trypan blue staining solution (2X) can also be directly added for staining.\u003c\/strong\u003e\u003cbr\u003e3. Counting:\u003cbr\u003eA small amount of stained cells was aspirated and counted on a blood count plate. Usually, if quantification is to be performed more accurately, at least 500 cells per cell sample, and the blue cells and the total number of cells are counted. The cell viability rate was calculated as follows:\u003cbr\u003e\u003cstrong\u003eCell viability = (total number of cells-number of blue cells)\/total number of cells × 100%\u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e         Trypan blue stained cell viability test kit (Trypan Blue Staining Cell Viability Assay Kit ), is developed by using normal healthy cells that can reject trypan blue, while cells that have lost cell membrane integrity can be stained with trypan blue. Trypan blue staining detects the integrity of the cell membrane,   It is generally considered that the cell membrane has lost its integrity, that is, the cell is considered dead. \u003cbr\u003e         After trypan blue staining, the cell survival rate can be quantified more accurately by counting directly under the microscope or counting after taking photos under the microscope. After trypan blue staining HeLa Please refer to Figure for cells 1 。 \u003cbr\u003e         Trypan blue staining only requires 3-5 It can be done in minutes, and it is very simple to operate. \u003cbr\u003e         This kit is sufficient to detect 100 Cell samples. \u003cbr\u003e\u003cstrong\u003e Product composition: \u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 95%; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.4641%; text-align: center;\"\u003e\u003cstrong\u003e Product Code \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 34.8484%; text-align: center;\"\u003e\u003cstrong\u003e Name \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 21.5512%; text-align: center;\"\u003e\u003cstrong\u003e100T\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 20.1311%; text-align: center;\"\u003e\u003cstrong\u003e500T\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.4641%; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 34.8484%; text-align: center;\"\u003e Trypan blue staining solution (2X)\u003c\/td\u003e\n\u003ctd style=\"width: 21.5512%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 20.1311%; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.4641%; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 34.8484%; text-align: center;\"\u003e Cell Resuspension \u003c\/td\u003e\n\u003ctd style=\"width: 21.5512%; text-align: center;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 20.1311%; text-align: center;\"\u003e500mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e1. A small amount of granular precipitate may appear when this product is stored for a long time. The precipitate can be fully dissolved in a 37 °C water bath for about 10 minutes. After the precipitate is completely dissolved, it can be used normally.\u003cbr\u003e2. Trypan blue is toxic to human body. Please be particularly careful when operating, and pay attention to effective protection to avoid direct contact with human body or inhalation.\u003cbr\u003e3. For your safety and health, please wear a laboratory coat and disposable gloves and masks.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStored at 2-8 ℃, the shelf life is 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"500T","offer_id":41293300301899,"sku":"abs50036-500T","price":80.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":43213556252747,"sku":"abs50036-100T","price":18.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/b1a2cc841c744e9ea4fea50dc66d41f4.png?v=1789740043"},{"product_id":"edu-594-cell-proliferation-detection-kit","title":"EdU-594 cell proliferation detection kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eBefore the trial guidelines\u003cbr\u003e1, the corresponding animal experiment, EdU suggested initial dosage of 50 mg\/kg (50 mu g\/g), dilute concentration is 0.5 1 mg\/mL. Are injected into mice weight 20 g, mg, 1 to 1 mg\/mL, need injection of 1 mL.\u003cbr\u003e2, for the cells, the recommended 10 microns, EdU work for in vitro culture cell, specific EdU use concentration, incubation time along with the samples and research purpose is different, can be adjusted appropriately.\u003cbr\u003e3, this kit can be carried out about 200 t of 6 orifice EdU experiments, about 2000 t96 orifice or paraffin section EdU experiment, experiment should be calculated according to the sample situation in the process of usage.\u003cbr\u003eI. Sample processing\u003cbr\u003e1, the animal experiment\u003cbr\u003e(1) EdU injection in animals\u003cbr\u003eIn mice, can according to the amount of 10-200 mg\/kg, the EdU with PBS mixed with a certain concentration, intraperitoneal injection, specific tissues or organs local injections or added to drinking water. The specific dosage is related to the type, weight and mode of use of animals, and can be referred to the relevant literature. Therefore, it is recommended to conduct a certain exploration of the use concentration of EdU for the first use, or directly use the concentration of 50mg\/kg for testing.\u003cbr\u003eInjection method: the experiment according to the customer, such as abdominal cavity injection, subcutaneous injection, intramuscular, tail intravenous injection, etc., among them with intraperitoneal injection.\u003cbr\u003eAfter 6 h or after the appropriate time was determined according to the specific experiment, the mice were quickly sacrificed, the desired tissue was removed, and frozen or paraffin sections were made according to routine procedures. EdU labeling can also be adjusted by referring to the relevant literature.\u003cbr\u003eSmall intestinal epithelial cells proliferate rapidly, and positive information can be detected 6 hours after EdU injection in adult mice, which can be used as a positive control for pre-experiments.\u003cbr\u003e(2) Slice processing\u003cbr\u003ePre-section treatment: It is best for tissues and organs to be washed to remove residual EdU in blood and tissues and reduce background.\u003cbr\u003eSection thickness: 3-10μm is appropriate, too thick section may affect the section background and staining efficiency.\u003cbr\u003eSlice post-processing:\u003cbr\u003eParaffin sections were deparaffinized twice with xylene for 15 min, once each with gradient ethanol (100%, 95%, 85%, 75%) for 5 min, and once with deionized water for elution.\u003cbr\u003eFrozen section processing: room temperature placed after 30 minutes, fixed 10 minutes, PBS cleaning three times, each time for 5 minutes.\u003cbr\u003e2. Cell experiments\u003cbr\u003e(1) cells EdU markers\u003cbr\u003ea.with cell complete medium and EdU mother liquor by 2000:1 the ratio of dilution, preparation of a moderate amount of 10 microns EdU medium;\u003cbr\u003eb.each hole adding suitable amount of 10 microns EdU medium 2 hours incubation, abandon culture medium (optimal incubation time for 1\/10 of the cell cycle) commonly, EdU incubation medium and EdU reaction liquid volume may refer to the table below;\u003cbr\u003ec.PBS cleaning cell 3 times, each time for 5 minutes.\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 94.6731%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e96-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e48-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e24-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e12-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e6-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e5.5cm capsule\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003eEdU medium\u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003eEdU reaction solution\u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e(2) cells fixed fully\u003cbr\u003ea. Cell fixative (PBS containing 4% paraformaldehyde) was added to each well and incubated at room temperature for 15 minutes. The fixative was discarded and washed with PBS for 3 times, 5 minutes each time.\u003cbr\u003eb. Each well was decolorized with an osmotic agent (0.5% TritonX-100 in PBS) and incubated for 15 minutes. PBS was washed three times for 5 min each.\u003cbr\u003e\u003cbr\u003eSecond, EdU response\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 50%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eConstituent\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eDosage\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e855μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e40μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e594 primary colour\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e5μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eReaction liquid volume\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e1000μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003eThe EdU reaction solution was prepared according to the ratio of PB: CU: AC: 594 chromogen = 855:40:100:5 (the reaction solution was prepared immediately).\u003cbr\u003eEach sample add 50-500 mu L EdU staining reaction liquid liquid evenly cover samples (reaction), 15 to 60 minutes at room temperature away from light incubation, abandon the dyeing liquid reaction, PBS wash 3 times, each time for 5 minutes.\u003cbr\u003e3. DAPI staining\u003cbr\u003eDAPI(100x) was diluted with PBS at a ratio of 1:100 to make ready-to-use DAPI staining solution. 50-500μL of DAPI staining solution was added to each sample and stained in the dark for 15 minutes, followed by three washes with PBS for 5 minutes each.\u003cbr\u003e4. Take pictures\u003cbr\u003eFluorescence microscope, confocal microscope or whole-film scanner were used to collect images. After staining, the slides were stored in a dark place at 4℃.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231114\/b72f553745434ba2afec62453f99c15d.png\" alt=\"\" width=\"753\" height=\"428\"\u003e\u003cbr\u003eAbove are stained images of 50mg\/kg labeled mice for 12 hours\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eEdU(5-ethynyl-2' -deoxyuridine), also known as 5-ethynyl-2' -deoxyuridine (5-ethynyl-2' -deoxyuridine), is a thymidine analogue with an alkynyl hydroxyl group that is rare in natural compounds and can replace thymidine (thymidine, 5-ethynyl-2' -deoxyuridine) in cell proliferation. thymidine (thymidine) is inserted into the replicating DNA molecule, the ethynyl group on EdU can react covalently with the fluorescently labeled small molecule Azide probe (Azide Alexa Fluor 488, etc.) through the catalysis of copper monovalent ion to form a stable triazole ring. \"It is called the Clickreaction, which allows for an efficient and rapid measurement of cell proliferation, especially the percentage of cells in S phase.\" With traditional immunofluorescence staining (BrdU) method compared with EdU only 1\/500 the size of the BrdU antibody, are more likely to spread within the cell, don't need strict sample degeneration (acid solution, pyrolysis, enzyme solution) processing, effectively avoid the damage samples, help in the overall level of tissue and organ observation of cell proliferation, It has higher sensitivity and faster detection speed.Note: AC is a powder, need to add 10mL pure water before use, fully dissolved before use, AC is easy to oxidation failure, after the refrigerator is stored at -20 degrees, if found discoloration, need to replace fresh reagent. In animal experiments, the mass concentration of 100mM EdU mother liquor corresponds to 25mg\/mL, which should be converted according to the dosage.\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 65.4028%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003eNO.\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003eSize\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003eStorage Temp.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e120mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e1.7g\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e4.4mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e594 Primary Colour\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e550μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e5\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e100mg\/mL EdU mother solution\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 12.6323%; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; text-align: center;\"\u003eDAPI（100×）\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; text-align: center;\"\u003e1.2mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003eNote: AC is a powder, need to add 10mL of pure water before use, fully dissolved before use, AC is easy to oxidation failure, after packaging and stored in the refrigerator -20 degrees, if found discoloration, need to replace fresh reagent. In animal experiments, the mass concentration of 100mM EdU mother liquor corresponds to 25mg\/mL, which should be converted according to the dosage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e-20 ° C, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"200T","offer_id":41293300432971,"sku":"abs50052-200T","price":440.0,"currency_code":"USD","in_stock":true}]},{"product_id":"edu-647-cell-proliferation-detection-kit","title":"EdU-647 cell proliferation detection kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eBefore the trial guidelines\u003cbr\u003e1, the corresponding animal experiment, EdU suggested initial dosage of 50 mg\/kg (50 mu g\/g), dilute concentration is 0.5 1 mg\/mL. Are injected into mice weight 20 g, mg, 1 to 1 mg\/mL, need injection of 1 mL.\u003cbr\u003e2, for the cells, the recommended 10 microns, EdU work for in vitro culture cell, specific EdU use concentration, incubation time along with the samples and research purpose is different, can be adjusted appropriately.\u003cbr\u003e3, this kit can be carried out about 200 t of 6 orifice EdU experiments, about 2000 t96 orifice or paraffin section EdU experiment, experiment should be calculated according to the sample situation in the process of usage.\u003cbr\u003eI. Sample processing\u003cbr\u003e1, the animal experiment\u003cbr\u003e(1) EdU injection in animals\u003cbr\u003eIn mice, can according to the amount of 10-200 mg\/kg, the EdU with PBS mixed with a certain concentration, intraperitoneal injection, specific tissues or organs local injections or added to drinking water. The specific dosage is related to the type, weight and mode of use of animals, and can be referred to the relevant literature. Therefore, it is recommended to conduct a certain exploration of the use concentration of EdU for the first use, or directly use the concentration of 50mg\/kg for testing.\u003cbr\u003eInjection method: the experiment according to the customer, such as abdominal cavity injection, subcutaneous injection, intramuscular, tail intravenous injection, etc., among them with intraperitoneal injection.\u003cbr\u003eAfter 6 h or after the appropriate time was determined according to the specific experiment, the mice were quickly sacrificed, the desired tissue was removed, and frozen or paraffin sections were made according to routine procedures. EdU labeling can also be adjusted by referring to the relevant literature.\u003cbr\u003eSmall intestinal epithelial cells proliferate rapidly, and positive information can be detected 6 hours after EdU injection in adult mice, which can be used as a positive control for pre-experiments.\u003cbr\u003e(2) Slice processing\u003cbr\u003ePre-section treatment: It is best for tissues and organs to be washed to remove residual EdU in blood and tissues and reduce background.\u003cbr\u003eSection thickness: 3-10μm is appropriate, too thick section may affect the section background and staining efficiency.\u003cbr\u003eSlice post-processing:\u003cbr\u003eParaffin sections were deparaffinized twice with xylene for 15 min, once each with gradient ethanol (100%, 95%, 85%, 75%) for 5 min, and once with deionized water for elution.\u003cbr\u003eFrozen section processing: room temperature placed after 30 minutes, fixed 10 minutes, PBS cleaning three times, each time for 5 minutes.\u003cbr\u003e2. Cell experiments\u003cbr\u003e(1) cells EdU markers\u003cbr\u003ea.with cell complete medium and EdU mother liquor by 2000:1 the ratio of dilution, preparation of a moderate amount of 10 microns EdU medium;\u003cbr\u003eb.each hole adding suitable amount of 10 microns EdU medium 2 hours incubation, abandon culture medium (optimal incubation time for 1\/10 of the cell cycle) commonly, EdU incubation medium and EdU reaction liquid volume may refer to the table below;\u003cbr\u003ec.PBS cleaning cell 3 times, each time for 5 minutes.\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 94.6731%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e96-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e48-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e24-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e12-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e6-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e5.5cm capsule\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003eEdU medium\u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003eEdU reaction solution\u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e(2) cells fixed fully\u003cbr\u003ea. Cell fixative (PBS containing 4% paraformaldehyde) was added to each well and incubated at room temperature for 15 minutes. The fixative was discarded and washed with PBS for 3 times, 5 minutes each time.\u003cbr\u003eb. Each well was decolorized with an osmotic agent (0.5% TritonX-100 in PBS) and incubated for 15 minutes. PBS was washed three times for 5 min each.\u003cbr\u003e\u003cbr\u003eSecond, EdU response\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 50%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eConstituent\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eDosage\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e855μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e40μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e647 primary colour\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e5μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eReaction liquid volume\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e1000μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003eThe EdU reaction solution was prepared according to the ratio of PB: CU: AC: 647 chromogen = 855:40:100:5 (the reaction solution was prepared immediately).\u003cbr\u003eEach sample add 50-500 mu L EdU staining reaction liquid liquid evenly cover samples (reaction), 15 to 60 minutes at room temperature away from light incubation, abandon the dyeing liquid reaction, PBS wash 3 times, each time for 5 minutes.\u003cbr\u003e3. DAPI staining\u003cbr\u003eDAPI(100x) was diluted with PBS at a ratio of 1:100 to make ready-to-use DAPI staining solution. 50-500μL of DAPI staining solution was added to each sample and stained in the dark for 15 minutes, followed by three washes with PBS for 5 minutes each.\u003cbr\u003e4. Take pictures\u003cbr\u003eFluorescence microscope, confocal microscope or whole-film scanner were used to collect images. After staining, the slides were stored in a dark place at 4℃.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231114\/b7c49145c81e4f3a9c43cf569b625e25.png\" alt=\"\" width=\"707\" height=\"414\"\u003e\u003cbr\u003eAbove are stained images of 50mg\/kg labeled mice for 12 hours\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eEdU(5-ethynyl-2' -deoxyuridine), also known as 5-ethynyl-2' -deoxyuridine (5-ethynyl-2' -deoxyuridine), is a thymidine analogue with an alkynyl hydroxyl group that is rare in natural compounds and can replace thymidine (thymidine, 5-ethynyl-2' -deoxyuridine) in cell proliferation. thymidine (thymidine) is inserted into the replicating DNA molecule, the ethynyl group on EdU can react covalently with the fluorescently labeled small molecule Azide probe (Azide Alexa Fluor 488, etc.) through the catalysis of copper monovalent ion to form a stable triazole ring. \"It is called the Clickreaction, which allows for an efficient and rapid measurement of cell proliferation, especially the percentage of cells in S phase.\" With traditional immunofluorescence staining (BrdU) method compared with EdU only 1\/500 the size of the BrdU antibody, are more likely to spread within the cell, don't need strict sample degeneration (acid solution, pyrolysis, enzyme solution) processing, effectively avoid the damage samples, help in the overall level of tissue and organ observation of cell proliferation, It has higher sensitivity and faster detection speed.Note: AC is a powder, need to add 10mL pure water before use, fully dissolved before use, AC is easy to oxidation failure, after the refrigerator is stored at -20 degrees, if found discoloration, need to replace fresh reagent. In animal experiments, the mass concentration of 100mM EdU mother liquor corresponds to 25mg\/mL, which should be converted according to the dosage.\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 65.4028%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003eNO.\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003eSize\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003eStorage Temp.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e120mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e1.7g\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e4.4mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e647 Primary Colour\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e550μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e5\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e100mg\/mL EdU mother solution\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 12.6323%; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; text-align: center;\"\u003eDAPI（100×）\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; text-align: center;\"\u003e1.2mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003eNote: AC is a powder, need to add 10mL of pure water before use, fully dissolved before use, AC is easy to oxidation failure, after packaging and stored in the refrigerator -20 degrees, if found discoloration, need to replace fresh reagent. In animal experiments, the mass concentration of 100mM EdU mother liquor corresponds to 25mg\/mL, which should be converted according to the dosage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e-20 ° C, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"200T","offer_id":41293300498507,"sku":"abs50053-200T","price":440.0,"currency_code":"USD","in_stock":true}]},{"product_id":"cell-titer-turbo-3d-luminescent-cell-viability-assay","title":"Cell Titer Turbo 3D Luminescent Cell Viability Assay","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003eReagent Preparation:\u003c\/strong\u003e\u003cbr\u003e1. Melting of reagents: Take out the reagents the day before the experiment and place them at 4 °C overnight to melt. The reagent can also be taken out on the day of the experiment and melted at room temperature, or melted in a 22 °C water bath, but it should be noted that the water temperature should not exceed 25 °C.\u003cbr\u003e2. Equilibrate to room temperature: If the reagent melts at non-room temperature conditions, it can be placed in a 22 °C water bath before use to ensure that the reagent is balanced to room temperature before being used for testing. Note: Generally, it takes about 10 minutes for 5mL packaging; The 50 mL packaging takes about 20 minutes.\u003cbr\u003e3. Mix well: Gently invert the solution 5 times before use to mix the solution evenly.\u003cbr\u003e\u003cstrong\u003eDetection Procedure:\u003c\/strong\u003e\u003cbr\u003e1. Use a 96-well plate suitable for chemiluminescence detection, and add microtissue containing cell culture medium to it.\u003cbr\u003e\u003cstrong\u003eNote: (1) The prepared sample volume and microtissue characteristics (e.g., size, quantity, culture days, etc.) should be optimized for experimental conditions; (2) The total ATP content of the sample should be kept below 10μM (determine the upper limit concentration of the linear range); (3) The multi-well plate must be compatible with the photometer used.\u003c\/strong\u003e\u003cbr\u003e2. Add test compounds to experimental wells and incubate according to your culture protocol.\u003cbr\u003e\u003cstrong\u003eNote: Ensure that the volume of sample and test compound is low enough to allow the addition of equal volumes of reagent and then mixing without inter-well contamination.\u003c\/strong\u003e\u003cbr\u003e3. Equilibrate the plate and its contents to room temperature (22-25 °C) for about 30 minutes.\u003cbr\u003e4. Add 3D Reagent equal to the volume of cell culture medium present in each well to the detection well (e.g., for a 96-well plate, add 100 μl of 3D Reagent to 100 μl of cell-containing culture medium).\u003cbr\u003e5. The contents were vigorously mixed for 5 minutes to induce cell lysis.\u003cbr\u003e\u003cstrong\u003eNote: Mixing is important for efficient ATP extraction from 3D microtissues.\u003c\/strong\u003e\u003cbr\u003e6. Allow the plate to incubate at room temperature for another 25 minutes to stabilize the luminescence signal.\u003cbr\u003e7. Record luminescence.\u003cbr\u003e\u003cstrong\u003eNote: The non-uniform luminescence signal within the plate may be caused by temperature gradients, non-uniform cell seeding, or edge effects in multi-well plates.\u003c\/strong\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e3D cell cultures are more physiologically relevant and better representative of tissues in vivo, allowing more accurate predictions of efficacy or toxicity of drug treatments. 3D cell models are increasingly being used to understand disease mechanisms and discover drug therapies. Compared with 2D culture, 3D cell culture can predict the efficacy or toxicity of drug treatment more accurately.\u003cbr\u003e3D Luminescent Cell Viability Assay Kit is a cell viability detection reagent based on luciferase system. It uses ATP-dependent luciferase-catalyzed luciferin luminescence reaction to measure intracellular ATP content through chemiluminescence signal to detect cell viability. The detection has wide linear range, high sensitivity and good stability.\u003cbr\u003eDue to the high-purity luciferin substrate, thermostable luciferase and optimized reaction reagents in this kit, this product has stronger cell lysis ability and can be used for cell viability analysis of 3D microtissue culture. This product is added to the cell culture to lyse the cell mass and release ATP, which can produce the reaction as shown in the figure and send out a stable light signal. The luminescence intensity is directly proportional to the amount of ATP, that is, the number of viable cells, within a certain range, so this product can quantitatively detect the number of viable cells.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231218\/595f7cf0567f40e98da696cd181a6cfc.png\" alt=\"\" width=\"493\" height=\"159\"\u003e\u003cbr\u003ePRODUCT FEATURES\u003cbr\u003eThe 3D Luminescent Cell Viability Assay has high detection sensitivity and wide linear range, and is compatible with small sample detection and high-throughput screening of large samples.\u003cbr\u003e1. Convenient and fast: The detection reagents provided in the kit are ready-to-use, with stable readings and fast detection speed. It only takes about 10 minutes to complete the detection;\u003cbr\u003e2. High sensitivity: It has a higher signal-to-noise ratio when applied to 3D microtissue detection;\u003cbr\u003e3. Fast: data can be recorded within 30 minutes or less after adding reagents;\u003cbr\u003e4. The luminescent signal is more stable: the half-life of the luminescent signal is ultra-stable, usually \u0026gt; 3 hours.\u003cbr\u003e5. Stronger lysis ability: The optimized reaction reagent has stronger cell lysis ability and is suitable for cell viability analysis of 3D microtissue culture;\u003cbr\u003e6. High throughput: It is compatible with the detection of a small amount of samples and the high throughput screening of a large number of samples.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Temperature: The reagent contains luciferase, and repeated freezing and thawing will affect its activity. It is recommended to store at-20 ℃ in the dark from light after dispensing. Reagents and cell samples should be balanced to room temperature before use to avoid the influence of enzyme catalytic effect;\u003cbr\u003e2. Chemical factors: The reaction rate and luminous intensity of luciferase are affected by the chemical environment. There are differences in luminescence intensity and attenuation rate among different types of media and serum. In addition, high drug content may interfere with the luciferase reaction, thus affecting the luminescence signal. It is recommended to set up control wells of cell culture medium containing drugs to eliminate the interference of solvents;\u003cbr\u003e3. Light sensitivity: This reagent is sensitive to light. If it is exposed to light during storage, it will accelerate the attenuation of luminous intensity. If the reagent is transferred from the original container, make sure to keep it protected from light;\u003cbr\u003e4. ATP contamination: It is recommended to wear masks and latex gloves during operation to avoid contact with surfaces and equipment that may be contaminated. Avoid inserting the tip of the gun tip into the vial multiple times during operation.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003ereagent preparation: \u003c\/strong\u003e\u003cbr\u003e1, reagent melting: The day before the experiment, the reagent was removed and placed at 4 ° C overnight to melt. It is also possible to remove the reagent on the day of the experiment and melt it at room temperature or place it in a 22℃ water bath, but it should be noted that the water temperature should not exceed 25℃. \u003cbr\u003e2, balance to room temperature: if the reagent melts under non-room temperature conditions, it can be placed in a 22 ° C water bath before use, to ensure that the reagent is balanced to room temperature before detection. Note: Generally, it takes about 10min for 5mL packaging; 50mL packaging takes about 20min. \u003cbr\u003e3, mixing: Before use, gently invert 5 times to mix the solution evenly. \u003cbr\u003e\u003cstrong\u003edetection steps: \u003c\/strong\u003e\u003cbr\u003e1, use a 96-well plate suitable for chemiluminescence detection, in which microtissue containing cell culture medium is added. \u003cbr\u003e\u003cstrong\u003eNote: (1) The preparation sample volume and microtissue characteristics (e.g., size, number, incubation days, etc.) should be optimized for the experimental conditions; (2) The total ATP content of the sample should be maintained at 10μ \u003cm of the upper limit linear range concentration multiwell plate must be compatible with photometer used.\u003e\u003cbr\u003e2, Add the test compound to the experimental well and incubate according to your culture protocol. \u003cbr\u003e\u003cstrong\u003eNote: Make sure the volume of the sample and test compound is low enough to allow the addition of an equal volume of reagent, followed by mixing, without interwell contamination. \u003c\/strong\u003e\u003cbr\u003e3, Equilibrate the plate and its contents to room temperature (22-25 ° C) for about 30 minutes. \u003cbr\u003e4, Add Cell Titer Turbo 3D Reagent equal to the volume of cell medium present in each well to the assay Wells (for example, for a 96-well plate, add 100μ l Cell Titer Turbo 3D Reagent was added to 100μ l in the medium containing the cells). \u003cbr\u003e5, mix the contents vigorously for 5 min to induce cell lysis. \u003cbr\u003e\u003cstrong\u003eNote: Mixing is important for efficient ATP extraction from 3D microtissues. \u003c\/strong\u003e\u003cbr\u003e6, Allow the plate to incubate at room temperature for an additional 25 minutes to stabilize the luminescence signal.  strong \u0026gt;\u003cbr\u003e7, record luminescence. \u003cbr\u003e\u003cstrong\u003eNote: The non-uniform luminescence signal in the plate may be caused by temperature gradients, uneven cell seeding, or edge effects in the multi-well plate. \u003c\/strong\u003e\u003c\/m\u003e\u003c\/strong\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Store below-10 °C and protect from light. In order to ensure the best performance, it is recommended to place it at-70 ℃ for long-term storage.\u003cbr\u003e2. This product can still remain stable after 6 cycles of repeated freezing and thawing.\u003cbr\u003e3. Sub-packaging may lead to the risk of ATP contamination, so avoid sub-packaging.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eIt is suitable for activity detection of bioactive factors, large-scale anti-tumor drug screening, cell proliferation, cytotoxicity test, etc.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50mL×2","offer_id":41293300596811,"sku":"abs50057-50mL×2","price":900.0,"currency_code":"USD","in_stock":true}]},{"product_id":"alamar-blue-cell-proliferation","title":"Alamar Blue cell proliferation and toxicity detection reagent","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eAppearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003esolution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e  Alamar Blue is a cell viability detection reagent, which contains an indicator with cell membrane permeability, non toxicity and weak blue fluorescence, namely resazurin. Resazurin has been cited as a reliable and reliable reagent since 1993. Alma blue is an effective and non-toxic alternative to MTT (thiazolam). Alma blue can quantitatively detect the proliferation of human, mammalian, bacterial, fungal and Mycoplasma cells, and can also be used for cytokine bioactivity, cell viability analysis, in vitro cytotoxicity determination and cell growth monitoring\u003cbr\u003e  The working principle of Alma blue is that resazurin is a redox indicator, which changes color according to cellular metabolic reduction. The oxidized resazurin is purple blue and basically non fluorescent, and its reduced product resorufin turns pink and highly fluorescent, and the fluorescence intensity is proportional to the number of living respiratory cells. By detecting the oxidation level during respiration, Alma blue is used as a direct indicator for quantitative detection of cell viability and toxicity. The color change of Alma blue can be detected by an ordinary spectrophotometer with a detection wavelength of 570nm and a reference wavelength of 600nm; The fluorescence change of Alma blue can be detected by a fluorophotometer, the excitation wavelength is between 530~560nm, and the emission wavelength is 590nm\u003cbr\u003e  Compared with trypan blue, TTC, MTT, MTS and other analytical methods, Alma blue is a single reagent, which can continuously and rapidly detect the proliferation of cells. Alma blue is non-toxic and harmless to cells, will not interfere with the electron transport chain, will not interfere with cell respiration or function, and will not affect the activity of antibody synthesis and secretion of cells. Therefore, it is suitable for continuous observation and further observation of the proliferation of the same batch of cells, and has the characteristics of simple operation and almost no interference with normal metabolism\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. The reduced Alamar Blue is very unstable in water or water-soluble buffer such as PBS, but very stable in culture medium. In order to determine the absorbance\/fluorescence values of the reduced Alamar Blue in a specific experiment, a 100% reduced Alamar Blue reagent needs to be prepared: Alamar Blue is mixed with cell culture medium in a ratio of 1:10, and high-pressure sterilization is sufficient for 15 minutes\u0026lt;2. Appropriate cell density can increase detection sensitivity. For 96 well plates, it is recommended to inoculate 100\u0026amp;mu per well; L cells, cell concentration range: adherent cells range from 100 to 10000 cells per well, suspended cells range from 2000 to 50000 cells per well, with culture medium as the blank control. For the 384 well plate, both cell concentration and inoculation amount were halved\u003cbr\u003e3. The two main variables that affect the cell response to Alamar Blue are incubation time and laying density. It is recommended to explore and optimize these two factors before conducting experiments. High cell density or prolonged incubation can lead to secondary reduction reactions, where the red product stops increasing and begins to decay, leading to a significant decrease in absorbance\/fluorescence levels and accompanied by the disappearance of red\u0026lt;4. Microbial contamination will reduce Alamar Blue. Therefore, using Alamar Blue detection on contaminated cells can cause erroneous results\u0026lt;5. Alamar Blue can be detected using a spectrophotometer or fluorescence meter, but it has high fluorescence sensitivity and small experimental error. Therefore, fluorescence detection is recommended\u003cfor your safety and health please wear laboratory clothes disposable gloves for operation.\u003e\u003c\/for\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplication\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCell proliferation and toxicity testing\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eProtected from light, 1 year\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"500T","offer_id":41683113050187,"sku":"abs47047610-500T","price":74.0,"currency_code":"USD","in_stock":true},{"title":"1000T","offer_id":41683113082955,"sku":"abs47047610-1000T","price":145.0,"currency_code":"USD","in_stock":true},{"title":"5000T","offer_id":41521807556683,"sku":"abs47047610-5000T","price":513.0,"currency_code":"USD","in_stock":true}]},{"product_id":"annexin-v-fitc-apoptosis-assay-kit","title":"Annexin V-FITC\/PI Apoptosis Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eOperating Instructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e(1) Centrifuge at low speed before use to avoid liquid accumulation in the tube cover and tube wall; \n(2) This reagent is only suitable for experiments, not for clinical testing; \n(3) PI (propidium iodide, propidium bromide) is toxic, can be absorbed through the skin, and has an irritating effect on the eyes. Wear gloves when using it; \n(4) Annexin V-FITC and PI are photosensitive substances, please pay attention to avoid light during operation. When handling and marking, do it in the dark as much as possible. During the incubation phase, the container is wrapped with aluminum foil or placed in a drawer. After the cells were labeled, they were observed with a microscope in a dark room; \n(5) The whole operation process should be as gentle as possible, do not blow the cells hard, and try to operate at 4 ℃ to avoid affecting the cell state. \n(6) In the last step of cell washing, please discard the supernatant as much as possible to avoid PBS residue, which may affect the experimental results. \n(7) In order to prevent fluorescence decay, flow cytometry detection should be performed within 1 hour. \n(8) If the PI staining time is too long, it may cause the detected apoptosis rate to be high. It is recommended to perform Annexin V-FITC staining first, and PI staining can be added at least 5 minutes before the machine. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAnnexin V-FITC apoptosis assay kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eProtocol\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAdherent cells need to be digested with 0.25% trypsin. Note that over-digestion can damage cells. 2% BSA can be added during digestion to prevent overdigestion. If digested with trypsin containing EDTA, be careful that EDTA must be completely removed: wash with 1 × PBS or 1 × binding buffer before labeling to remove EDTA to prevent residual EDTA from chelating with Ca2 + and affecting the binding of Annexin V. \n(1) Dilute 10 × Binding Buffer into 1 × Binding Buffer with deionized water; \n(2) Cell collection: suspension cell collection: centrifugation for 5 minutes; Adherent cells: After digestion and collection with trypsin without EDTA (Note: the trypsin digestion time should not be too long, otherwise it will affect the binding of phosphatidylserine and Annexin V-FITC on the cell membrane), centrifuge at 2000 rpm at room temperature for 5 ~ 10 minutes to collect the cells; \n(3) Cell washing: Resuspend the cells once with pre-cooled 1 × PBS (4 ℃), centrifuge at 2000 rpm for 5-10 minutes, and discard the supernatant to wash the cells; \n(4) 300 μL of 1 × Binding Buffer suspension cells were added; \n(5) Annexin V-FITC labeling: Add 5μL of Annexin V-FITC and mix well, protect from light, and incubate at room temperature for 15 minutes; \n(6) PI labeling: Add 5μL of PI staining 5 minutes before loading the machine. \n(7) Before loading, add 200 μL of 1 × Binding Buffer. \nNote: The recommended number of cells per reaction system is 1 × 10 \u003csup\u003e4\u003c\/sup\u003eTo 1 × 10\u003csup\u003e6\u003c\/sup\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e In normal cells, phosphatidylserine is only distributed on the inner side of the cell membrane lipid bilayer. At the earliest stage of cell apoptosis, membrane phosphatidylserine ( PS ) from the inside to the outside of the lipid membrane, this change is earlier than cell shrinkage, chromatin condensation, DNA Apoptotic phenomena such as fragmentation and increased permeability of cell membrane. Annexin V It is a phospholipid-binding protein with high affinity for phosphatidylserine, so it can bind to the cell membrane of early apoptotic cells through the exposed phosphatidylserine on the outside of the cell. Hence Annexin V It is used as one of the sensitive indicators to detect early apoptosis of cells. Propidium iodide ( Propidium Iodide, PI ) is a nucleic acid dye ,  It cannot penetrate the intact cell membrane, but cells and dead cells in the middle and late stages of apoptosis due to the increase of cell membrane permeability, PI  Can penetrate the cell membrane and make the nucleus red. Thus will Annexin V  With PI  With matching, cells in different apoptotic stages can be distinguished. \u003cbr\u003e\u003cstrong\u003e Product composition: \u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 95%; margin-left: auto; margin-right: auto; height: 88px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.238%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e Name \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 18.5246%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e25T\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 21.9144%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e50T\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 19.295%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e100T\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.238%; text-align: center; height: 22px;\"\u003eAnnexinV-FITC\u003c\/td\u003e\n\u003ctd style=\"width: 18.5246%; text-align: center; height: 22px;\"\u003e125μL\u003c\/td\u003e\n\u003ctd style=\"width: 21.9144%; text-align: center; height: 22px;\"\u003e250μL\u003c\/td\u003e\n\u003ctd style=\"width: 19.295%; text-align: center; height: 22px;\"\u003e500μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.238%; text-align: center; height: 22px;\"\u003e10×Binding Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 18.5246%; text-align: center; height: 22px;\"\u003e1.5mL\u003c\/td\u003e\n\u003ctd style=\"width: 21.9144%; text-align: center; height: 22px;\"\u003e2.5mL\u003c\/td\u003e\n\u003ctd style=\"width: 19.295%; text-align: center; height: 22px;\"\u003e6mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 32.238%; text-align: center; height: 22px;\"\u003ePropidium Iodide\u003c\/td\u003e\n\u003ctd style=\"width: 18.5246%; text-align: center; height: 22px;\"\u003e125μL\u003c\/td\u003e\n\u003ctd style=\"width: 21.9144%; text-align: center; height: 22px;\"\u003e250μL\u003c\/td\u003e\n\u003ctd style=\"width: 19.295%; text-align: center; height: 22px;\"\u003e500μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e4 °C for 6 months; Avoid repeated freezing and thawing.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"25T","offer_id":41683110821963,"sku":"abs50001-25T","price":143.0,"currency_code":"USD","in_stock":true},{"title":"50T","offer_id":41683110854731,"sku":"abs50001-50T","price":206.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41521807687755,"sku":"abs50001-100T","price":317.0,"currency_code":"USD","in_stock":true}]},{"product_id":"cell-counting-kit","title":"CCK-8 Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCCK8,CCK 8,Cell Counting Kit-8\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eCell Counting Kit 8, referred to as CCK-8 kit for short, is a highly sensitive, non-radioactive colorimetric detection method used to determine the number of living cells in cell proliferation or toxicity experiments.\u003c\/p\u003e\n\u003cdiv\u003e \u003c\/div\u003e\n\u003cdiv\u003eCCK-8 is an upgraded product of MTT. CCK-8 solution can be directly added to cell samples without pre-preparing various ingredients. It has rapid detection and very low toxicity. CCK-8 is based on the water-soluble tetrazolium salt WST-8. The working principle is: WST-8 can be reduced by dehydrogenase in mitochondria to produce orange-yellow formazan dye in the presence of electron coupling reagent. The formazan dye is able to dissolve in tissue culture medium in proportion to the number of viable cells. Through colorimetry, the number of viable cells can be dynamically quantified, so as to detect cell proliferation or drug toxicity. This kit is usually used for cycle and apoptosis detection of adherent or suspended cells. If it is used in tissue, the tissue needs to be digested into a single-cell state, and then it can be tested.\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplication\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis product is restricted to scientific purposes only.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eIt is effective for one year when stored at 4 ℃ in the dark, and for two years when stored at-20 ℃ in the dark.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"500T","offer_id":43115013406795,"sku":"abs50003-500T","price":107.0,"currency_code":"USD","in_stock":true},{"title":"5000T","offer_id":41683118751819,"sku":"abs50003-5000T","price":714.0,"currency_code":"USD","in_stock":true}]},{"product_id":"cell-cycle-assay-kit","title":"Cell Cycle Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eSelf-provided consumables and equipment:\u003cbr\u003e1 ml pipette\u003cbr\u003e100-200 μl pipette\u003cbr\u003eRotary socket mixer\u003cbr\u003eFlow cytometry\u003cbr\u003ePBS\u003cbr\u003e75% ethanol\u003cbr\u003e\u003cbr\u003eApplicable experiment and operation process:\u003cbr\u003e1. Cell preparation Adherent cells: Discard the cell culture medium, digest with trypsin, prepare a single cell suspension, centrifuge at 1000 rpm for 5 minutes, and discard the supernatant. The pellet was resuspended with 1 ml of pre-cooled PBS. Centrifuge again at 1000 rpm for 5 minutes and discard the supernatant. Suspended cells: The cell suspension was collected, centrifuged at 1000 rpm for 5 minutes, and the supernatant was discarded. The pellet was resuspended with 1 ml of pre-cooled PBS. Centrifuge again at 1000 rpm for 5 minutes and discard the supernatant. Histiocytes: After cutting the tissue pieces into the smallest possible pieces with scissors, digested with 0.25% trypsin for 0.5-1 hour. Filtering through a 200-400 mesh screen to obtain a single cell suspension. Centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and resuspend the pellet with 1 ml of pre-cooled PBS. Centrifuge again at 1000 rpm for 5 minutes and discard the supernatant. Note that for the last centrifugation, discard the supernatant and leave 50 μl of the supernatant, vortex and mix well.\u003cbr\u003e2. Cell fixation The cell pellet is gently mixed with 1 ml-20 °C pre-cooled 75% ethanol, and fixed at 4 °C for more than 2 hours or overnight. Then, centrifuge at 1000 rpm for 5 minutes, gently discard the supernatant, resuspend the pellet with 1 ml of pre-cooled PBS, centrifuge at 1000 rpm for 5 minutes, discard the supernatant.\u003cbr\u003e3. Staining PI staining working solution configuration: Add 25μl PI staining solution (solution A) and 10μl RNase A (solution B) to 0.5 ml staining buffer solution (solution C), mix well and set aside. Add 0.5 ml of the prepared PI staining working solution to each cell sample, and gently mix and resuspend the cells. Incubate at 37 ℃ for 30 minutes in the dark from light, and detect directly by flow cytometry (it is preferable to complete within 5 hours). The excitation wavelength was 488 nm and red fluorescence was detected. Note: Not more than 1 × 10 cells per test\u003csup\u003e6\u003c\/sup\u003eOne.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCell cycle and apoptosis detection kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eThis product is based on Propidium iodide (PI) staining method to analyze cell cycle and apoptosis. Propidium iodide is a double-stranded DNA dye. Its intercalation in double-stranded DNA can produce fluorescence. The intensity of fluorescence is directly proportional to the amount of double-stranded DNA.\u003cbr\u003eIn the normal cell cycle, there is one set of chromosomes in G0 and G1 phases, two sets of chromosomes in G2 and M phases cells, and the S phase is in between. After PI staining, the fluorescence intensity of cells in different cell cycles is different. Assuming that the fluorescence intensity of cells in G0 and G1 phases is 1, then the fluorescence intensity of cells in G2 and M phases is 2, and the cells in S phase are between 1 and 2.\u003cbr\u003eDuring apoptosis, due to nucleus shrinkage and DNA fragmentation, DNA fragments will be lost from the perforated cell membrane during staining. When detected by flow cytometry, the fluorescence intensity is less than 1, that is, Sub-G1 peak or apoptotic cell peak.\u003cbr\u003eApoptosis can also be detected by observing changes in cell light scattering by flow cytometry. In the early stage of apoptosis, chromatin shrinks, cell density increases, and forward angle light dispersion decreases significantly. In the later stage of apoptosis, cells produce apoptotic bodies, and both forward angular light scattering and lateral angular light scattering are significantly reduced.\u003cbr\u003e\u003cbr\u003eProduct Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 47.6576%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.7955%; text-align: center;\"\u003eserial number\u003c\/td\u003e\n\u003ctd style=\"width: 19.0183%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 11.0599%; text-align: center;\"\u003e50T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.7955%; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 19.0183%; text-align: center;\"\u003ePropidium Iodide (20 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 11.0599%; text-align: center;\"\u003e1.25 ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.7955%; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 19.0183%; text-align: center;\"\u003eRNase A (50 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 11.0599%; text-align: center;\"\u003e500 μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.7955%; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 19.0183%; text-align: center;\"\u003eStaining Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 11.0599%; text-align: center;\"\u003e25ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. PI is toxic. Pay attention to protection during operation, protect eyes, avoid inhalation, and wear disposable gloves.\u003cbr\u003e2. PI has quenching phenomenon, so avoid light during storage and use.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at-20 ℃ in the dark from light, shelf life is 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis product is used for the detection of cultured adherent cells or suspended cells, and can also be used for the detection of tissue cells.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50T","offer_id":41518126694475,"sku":"abs50005-50T","price":100.0,"currency_code":"USD","in_stock":true}]},{"product_id":"annexin-v-egfp-pi-apoptosis-analysis-kit","title":"Annexin V-EGFP\/PI Apoptosis analysis kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Adjust the cell density to about 1 × 10\u003csup\u003e6\u003c\/sup\u003eCells\/ml. 2. Remove 0.2 ml of cell suspension from a cell culture flask (2 × 10\u003csup\u003e5\u003c\/sup\u003eCells) were transferred into a clean centrifuge tube. 3. Centrifuge at 1000 g at room temperature for 5 minutes to remove the medium. 4. The cell pellet was gently resuspended with 0.2 ml of pre-cooled PBS solution. 5. Centrifuge at 1000 g for 5 minutes at room temperature to remove PBS. 6. The cell pellet was gently resuspended with 0.2 ml of pre-cooled binding buffer (component C. Binding Buffer). 7. Add 5μl Annexin V-EGFP (component A) and 5μl PI dye solution (component B), and gently mix well. 8. Incubate at room temperature (18-24 °C) for 10-15 minutes in the dark. 9. Alternatively, replenish the volume to 0.4 ml with binding buffer or PBS. 10. Place the sample on ice and protect it from light, and test it as soon as possible. 11. Use flow cytometry or fluorescence microscope for detection and analysis.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAnnexin V-EGFP\/PI Apoptosis Analysis Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eAnnexin V-EGFP\/PI cell apoptosis detection kit uses EGFP-labeled Annexin V as a probe to detect the occurrence of early cell apoptosis.\u003c\/p\u003e\n\u003cp\u003eThe detection principle is: in normal living cells, phosphotidylserine (PS) is located on the inside of the cell membrane, but in early apoptotic cells, PS flips from the inside of the cell membrane to the surface of the cell membrane and is exposed to the extracellular environment. Annexin-V (Annexin-V) is a Ca2 +-dependent phospholipid binding protein with a molecular weight of 35-36KD, which can bind to PS with high affinity. It can bind to the cell membrane of early apoptotic cells through phosphatidylserine exposed on the outside of the cell.\u003c\/p\u003e\n\u003cdiv\u003eIn addition, Propidium Iodide (PI) is also provided in this kit to distinguish surviving early cells from necrotic or late apoptotic cells. PI is a nucleic acid dye that cannot penetrate the intact cell membrane of normal cells or early apoptotic cells, but can penetrate the cell membrane of late apoptotic and necrotic cells to make the nucleus red. Therefore, when Annexin V is used in combination with PI, PI is excluded from viable cells (Annexin V-\/PI-) and early apoptotic cells (Annexin V +\/PI-), while late apoptotic cells and necrotic cells are stained double positive by EGFP and PI combination (Annexin V +\/PI +).\u003c\/div\u003e\n\u003cdiv\u003eProduct Components:\u003c\/div\u003e\n\u003cdiv\u003e\u003ctable border=\"0\" cellpadding=\"0\" cellspacing=\"0\" width=\"565\" height=\"112\" style=\"border-collapse:collapse;width:282.52pt;\"\u003e\u003ctbody\u003e\n\u003ctr height=\"28\" style=\"height:14.00pt;\"\u003e\n\u003ctd class=\"et3\" height=\"28\" width=\"82\" style=\"height:14.00pt;width:41.00pt;\"\u003eserial number\u003c\/td\u003e\n\u003ctd class=\"et4\" width=\"207\" style=\"width:103.50pt;\"\u003eName\u003c\/td\u003e\n\u003ctd class=\"et4\" width=\"145\" style=\"width:72.50pt;\"\u003e20T\u003c\/td\u003e\n\u003ctd class=\"et4\" width=\"131\" style=\"width:65.50pt;\"\u003e50T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr height=\"28\" style=\"height:14.00pt;\"\u003e\n\u003ctd class=\"et4\" height=\"28\" style=\"height:14.00pt;\"\u003eA\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003eAnnexin V-EGFP\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003e100ul\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003e250ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr height=\"28\" style=\"height:14.00pt;\"\u003e\n\u003ctd class=\"et3\" height=\"28\" style=\"height:14.00pt;\"\u003eB\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003ePI\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003e100ul\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003e250ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr height=\"28\" style=\"height:14.00pt;\"\u003e\n\u003ctd class=\"et4\" height=\"28\" style=\"height:14.00pt;\"\u003eC\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003eBinding Buffer\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003e5ml\u003c\/td\u003e\n\u003ctd class=\"et4\"\u003e10ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStored at 2-8 ℃ in the dark from light, the validity period is 6 months. Store at-20 ℃ in the dark from light, shelf life for 12 months or more, avoid repeated freezing and thawing.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"20T","offer_id":41683111018571,"sku":"abs50006-20T","price":143.0,"currency_code":"USD","in_stock":true},{"title":"50T","offer_id":41521809424459,"sku":"abs50006-50T","price":206.0,"currency_code":"USD","in_stock":true}]},{"product_id":"annexin-v-pe-7-aad-apoptosis-analysis-kit","title":"Annexin V-PE\/7-AAD apoptosis analysis kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Instrument parameter adjustment\u003cbr\u003e1. Collect 1 × 10\u003csup\u003e6\u003c\/sup\u003e-3×10\u003csup\u003e6\u003c\/sup\u003eThe cells were washed twice by centrifugation with pre-cooled PBS and the supernatant was discarded.\u003cbr\u003e2. Add 500uL of Apoptosis Positive Control Solution to resuspend and incubate on ice for 30 minutes.\u003cbr\u003e3. Wash by centrifugation with pre-cooled PBS and discard the supernatant.\u003cbr\u003e4. Add an appropriate amount of pre-cooled 1 × Binding Buffer to resuspend, and add the same number of untreated viable cells to mix with it. Add pre-cooled 1 × Binding Buffer to replenish to 1.5 mL, and divide it into three tubes equally, of which one tube is a blank control tube and two tubes are single dye tubes.\u003cbr\u003e5. Add 5 uL of Annexin V-PE or 10 uL of 7-AAD to the single stain tube and incubate for 5 minutes at room temperature and protect from light.\u003cbr\u003e6. On the flow cytometer, the voltages of FSC, SSC and fluorescence channel were adjusted with a blank tube, and under this voltage condition, the fluorescence was adjusted with a single staining tube\u003cbr\u003eCompensation of channels.\u003cbr\u003e\u003cstrong\u003eNote:\u003c\/strong\u003eCertain processes of treating adherent cells as single cells cause cell membrane damage, resulting in false positives for Annexin V. Therefore, optimization is needed. It is recommended to treat adherent cells with enzymes that are more gentle on cells, such as Accutase.\u003cbr\u003eThe adherent cells can be cultured on a suitable vessel, such as a sterile glass slide, and stained with suitably modified methods. However, this method has not been tested in our laboratory.\u003cbr\u003e\u003cbr\u003e2. Sample testing\u003cbr\u003e1. Induce apoptosis according to the experimental protocol.\u003cbr\u003e2. Centrifuge washing with pre-cooled PBS and collect 1-10 × 10\u003csup\u003e5\u003c\/sup\u003eCells include cells in the culture supernatant. Dilute 5 × Binding Buffer into 1 × working solution with double distilled water, and take 500 uL of 1 × Binding Buffer to resuspend the cells.\u003cbr\u003e3. Add 5 uL Annexin V-PE and 10 uL 7-AAD per tube.\u003cbr\u003e4. After gently vortex and mix well, incubate at room temperature and protect from light for 5 minutes.\u003cbr\u003e5. Get on the computer for analysis.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAnnexin V-PE\/7-AAD Apoptosis Analysis Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eAnnexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. PS is present in the inner lining of the plasma membrane of normal cells, but in the early stage of apoptosis, membrane asymmetry is lost and PS translocates to the cell surface. Fluorescently Labeled Annexin V\u003cbr\u003eCan specifically bind to it, indicating that the cell is an apoptotic cell. This kit takes only 15 minutes to stain, which is quick and easy.\u003c\/p\u003e\n\u003cp\u003eKit Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 73.4507%; margin: 0 auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.3148%; text-align: center;\" width=\"160\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 14.2415%; text-align: center;\" width=\"64\"\u003e25T\u003c\/td\u003e\n\u003ctd style=\"width: 12.6612%; text-align: center;\" width=\"68\"\u003e100T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.3148%;\" width=\"160\"\u003eAnnexin V-PE\u003c\/td\u003e\n\u003ctd style=\"width: 14.2415%;\"\u003e150uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6612%;\"\u003e500uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.3148%;\" width=\"160\"\u003e7-AAD\u003c\/td\u003e\n\u003ctd style=\"width: 14.2415%;\"\u003e300uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6612%;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.3148%;\" width=\"160\"\u003e5 × Binding buffer\u003c\/td\u003e\n\u003ctd style=\"width: 14.2415%;\"\u003e5mL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6612%;\"\u003e15mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.3148%;\" width=\"160\"\u003eApoptosis Positive Control Solution\u003c\/td\u003e\n\u003ctd style=\"width: 14.2415%;\"\u003e5mL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6612%;\"\u003e5mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Please read the instructions carefully before using this product. This product is for scientific research only, not diagnostic.\u003cbr\u003e2. For your safety and health, please wear experimental protective clothing, gloves, masks and other necessary protective equipment.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at 2-8 °C, do not freeze, shelf life 1 year.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"25T","offer_id":41683111051339,"sku":"abs50007-25T","price":143.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41521809522763,"sku":"abs50007-100T","price":254.0,"currency_code":"USD","in_stock":true}]},{"product_id":"annexin-v-apc-7-aad-apoptosis-analysis-kit","title":"Annexin V-APC\/7-AAD apoptosis analysis kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Instrument parameter adjustment\u003cbr\u003e1. Collect 1 × 10\u003csup\u003e6\u003c\/sup\u003e-3×10\u003csup\u003e6\u003c\/sup\u003eThe cells were washed twice by centrifugation with pre-cooled PBS and the supernatant was discarded.\u003cbr\u003e2. Add 500uL of Apoptosis Positive Control Solution to resuspend and incubate on ice for 30 minutes.\u003cbr\u003e3. Wash by centrifugation with pre-cooled PBS and discard the supernatant.\u003cbr\u003e4. Add an appropriate amount of pre-cooled 1 × Binding Buffer to resuspend, and add the same number of untreated viable cells to mix with it. Add pre-cooled 1 × Binding Buffer to replenish to 1.5 mL, and divide it into three tubes equally, of which one tube is a blank control tube and two tubes are single dye tubes.\u003cbr\u003e5. Add 5 uL of Annexin V-APC or 10 uL of 7-AAD to the single stain tube and incubate for 5 minutes at room temperature and protect from light.\u003cbr\u003e6. On the flow cytometer, the voltages of FSC, SSC and fluorescence channel were adjusted with a blank tube, and under this voltage condition, the fluorescence was adjusted with a single staining tube\u003cbr\u003eCompensation of channels.\u003cbr\u003e\u003cstrong\u003eNote:\u003c\/strong\u003eCertain processes of treating adherent cells as single cells cause cell membrane damage, resulting in false positives for Annexin V. Therefore, optimization is needed. It is recommended to treat adherent cells with enzymes that are more gentle on cells, such as Accutase.\u003cbr\u003e\u003cbr\u003e2. Sample testing\u003cbr\u003e1. Induce apoptosis according to the experimental protocol.\u003cbr\u003e2. Centrifuge washing with pre-cooled PBS and collect 1-10 × 10\u003csup\u003e5\u003c\/sup\u003eCells (including cells in the culture supernatant). Dilute 5 × Binding Buffer into 1 × working solution with double distilled water, and take 500 uL of 1 × Binding Buffer to resuspend the cells.\u003cbr\u003e3. Add 5 uL of Annexin V-APC and 10 uL of 7-AAD per tube.\u003cbr\u003e4. After gently vortex and mix well, incubate at room temperature and protect from light for 5 minutes.\u003cbr\u003e5. Get on the computer for analysis.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAnnexin V-APC\/7-AAD Apoptosis Analysis Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eAnnexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. PS is present in the inner lining of the plasma membrane of normal cells, but in the early stage of apoptosis, membrane asymmetry is lost and PS translocates to the cell surface. Fluorescently Labeled Annexin V\u003cbr\u003eCan specifically bind to it, indicating that the cell is an apoptotic cell. This kit takes only 15 minutes to stain, which is quick and easy.\u003c\/p\u003e\n\u003cp\u003eKit Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 73.4506%; margin: 0 auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.5676%; text-align: center;\" width=\"160\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 12.8999%; text-align: center;\" width=\"64\"\u003e25T\u003c\/td\u003e\n\u003ctd style=\"width: 12.8829%; text-align: center;\" width=\"68\"\u003e100T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.5676%;\" width=\"160\"\u003eAnnexin V-APC\u003c\/td\u003e\n\u003ctd style=\"width: 12.8999%;\"\u003e150uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.8829%;\"\u003e500uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.5676%;\" width=\"160\"\u003e7-AAD\u003c\/td\u003e\n\u003ctd style=\"width: 12.8999%;\"\u003e300uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.8829%;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.5676%;\" width=\"160\"\u003e5 × Binding buffer\u003c\/td\u003e\n\u003ctd style=\"width: 12.8999%;\"\u003e5mL\u003c\/td\u003e\n\u003ctd style=\"width: 12.8829%;\"\u003e15mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.5676%;\" width=\"160\"\u003eApoptosis Positive Control Solution\u003c\/td\u003e\n\u003ctd style=\"width: 12.8999%;\"\u003e5mL\u003c\/td\u003e\n\u003ctd style=\"width: 12.8829%;\"\u003e5mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Please read the instructions carefully before using this product. This product is for scientific research only, not diagnostic.\u003cbr\u003e2. For your safety and health, please wear experimental protective clothing, gloves, masks and other necessary protective equipment.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at 2-8 °C, do not freeze, shelf life 1 year.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"25T","offer_id":41683111084107,"sku":"abs50008-25T","price":143.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41521810669643,"sku":"abs50008-100T","price":317.0,"currency_code":"USD","in_stock":true}]},{"product_id":"annexin-v-apc-pi-apoptosis-analysis-kit","title":"Annexin V-APC\/PI apoptosis analysis kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e1. Self-prepared instruments and reagents:\u003c\/strong\u003e\u003cbr\u003e1. Reagent: PBS (abs962); EDTA-free trypsin digestion (abs47014937, abs47048223)\u003cbr\u003e2. Consumables and instruments: 1.5 mL centrifuge tube (sterile and enzyme-free) (abs7119), flow cytometer, fluorescence microscope, low-speed centrifuge, micropipette.\u003cbr\u003e\u003cstrong\u003e2. Operation steps:\u003c\/strong\u003e\u003cbr\u003e\u003cstrong\u003eFor streaming operation detection\u003c\/strong\u003e\u003cbr\u003e1. Cell collection\u003cbr\u003e(1) Suspended cells: 300 × g centrifugal force, centrifugation for 5 min to collect;\u003cbr\u003e(2) Adherent cells: Adherent cells are collected by trypsin digestion without EDTA (Note: the trypsin digestion time should not be too long, otherwise it will easily cause false positives);\u003cbr\u003e2. Wash the cells twice with PBS (300 × g, centrifuge for 5 min) and collect 1-5 × 10\u003csup\u003e5\u003c\/sup\u003eCells;\u003cbr\u003e3. Add 500μL of Binding Buffer and gently blow to form a single cell suspension;\u003cbr\u003e4. Add 5μL Annexin V-APC and 5μL Propidium Iodide, and gently mix;\u003cbr\u003e5. React for 5-10 min at room temperature and in the dark from light;\u003cbr\u003e6. Please perform flow cytometer observation and detection within 1 hour.\u003cbr\u003e7. Flow cytometry analysis\u003cbr\u003e(1) Detection by flow cytometry, fluorescence channel (FL4) of Annexin V-APC (Ex = 633 nm, Em = 660 nm); PI red fluorescence (Ex = 488 nm, Em ≥ 630 nm) was detected by PE channels (FL2 or FL3).\u003cbr\u003e(2) Fluorescence compensation adjustment: Using induction-treated apoptotic cells as a control, fluorescence compensation adjustment was performed to remove spectral overlap and set the position of cross gates.\u003cbr\u003e\u003cstrong\u003eFor fluorescence microscopy detection\u003c\/strong\u003e\u003cbr\u003e1. Select a 24-well plate or a 48-well plate for cell culture and treatment;\u003cbr\u003e2. Before staining, use a multi-well plate centrifuge 300 × g for 5 min;\u003cbr\u003e3. Remove the cell culture medium and add PBS to wash once;\u003cbr\u003e4. According to the number of samples, calculate and prepare the staining working solution: in a 1.5 mL EP tube, add 500μL Binding Buffer, 5μL Annexin V-APC and 5μL Propidium Iodide, and mix well (take 500μL as an example);\u003cbr\u003e5. Add the staining working solution to the well plate (500μL per well for the 12-well plate, 200μL per well for the 24-well plate, 100μL per well for the 96-well plate), and incubate at room temperature in the dark for 20 minutes;\u003cbr\u003e6. Observe immediately under a fluorescence microscope.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAnnexin V-APC\/PI Apoptosis Analysis Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eIn normal cells, phosphatidylserine (PS) is only distributed on the inside of the lipid bilayer of the cell membrane, while in the early stage of apoptosis, phosphatidylserine (PS) in the cell membrane turns from the inside to the outside of the lipid membrane. Annexin V is a Ca2 +-dependent phospholipid binding protein with a molecular weight of 35-36 kD. It has a high affinity for phosphatidylserine, so it can bind to the cell membrane of early apoptotic cells through the exposed phosphatidylserine on the outside of the cell. Therefore, Annexin V is one of the sensitive indicators to detect early apoptosis. Annexin V was labeled with fluorescein APC, and the labeled Annexin V was used as a fluorescent probe to detect the occurrence of apoptosis by fluorescence microscope or flow cytometry.\u003cbr\u003ePropidium Iodide (PI) is a nucleic acid dye that cannot penetrate the intact cell membrane, but for cells in the middle and late stages of apoptosis and dead cells, PI can penetrate the cell membrane and make the nucleus red. Therefore, matching Annexin V-APC with PI can distinguish cells in different apoptotic stages.\u003cbr\u003eThis kit can be applied to the apoptosis detection of cultured cells (not recommended for the detection of tissue samples).\u003cbr\u003eProduct composition:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 80.3563%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 18.2656%; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 13.2911%; text-align: center;\"\u003e20T\u003c\/td\u003e\n\u003ctd style=\"width: 12.9085%; text-align: center;\"\u003e50T\u003c\/td\u003e\n\u003ctd style=\"width: 12.5258%; text-align: center;\"\u003e100T\u003c\/td\u003e\n\u003ctd style=\"width: 14.6941%; text-align: center;\"\u003eSave\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 18.2656%; text-align: center;\"\u003eAnnexin V-APC\u003c\/td\u003e\n\u003ctd style=\"width: 13.2911%; text-align: center;\"\u003e100uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.9085%; text-align: center;\"\u003e250uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5258%; text-align: center;\"\u003e500uL\u003c\/td\u003e\n\u003ctd style=\"width: 14.6941%; text-align: center;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 18.2656%; text-align: center;\"\u003ePropidium Iodide\u003c\/td\u003e\n\u003ctd style=\"width: 13.2911%; text-align: center;\"\u003e100uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.9085%; text-align: center;\"\u003e250uL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5258%; text-align: center;\"\u003e500uL\u003c\/td\u003e\n\u003ctd style=\"width: 14.6941%; text-align: center;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 18.2656%; text-align: center;\"\u003eBinding Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 13.2911%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 12.9085%; text-align: center;\"\u003e25mL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5258%; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 14.6941%; text-align: center;\"\u003e2-8℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Please centrifuge and collect the solution before taking the trace reagent.\u003cbr\u003e2. Annexin V-APC, Propidium Iodide (PI) is stored and used in the dark from light.\u003cbr\u003e3. Propidium Iodide (PI) is toxic, please wear gloves when operating.\u003cbr\u003e4. This kit is suitable for detecting living cells. When detected by flow cytometry, the number of cells should not be less than 1 × 10\u003csup\u003e5\u003c\/sup\u003eUnits\/mL, not recommended for testing tissue samples.\u003cbr\u003e5. Adherent cells need to be digested with trypsin without EDTA. If digested improperly, it may cause false positives, and using a cell scraper will cause cells to adhere and form clusters, which will affect the detection. Trypsinized cells can be stored in PBS containing 2% (w\/v) BSA to prevent further damage.\u003cbr\u003e6. The fluorescence may be quenched after cell fixation. Please do not fix the sample.\u003cbr\u003e7. Due to different types of detected cells, types of apoptosis inducers, detection instruments used, and different setting parameters, it is recommended that cells treated with apoptosis induction should be used as controls for each detection to adjust fluorescence compensation.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eKeep as concentrated solution. Store at 4°C and protected from prolonged exposure to light. Do not freeze.The validity period is 6 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"25T","offer_id":43115016650827,"sku":"abs50009-25T","price":317.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":43115016683595,"sku":"abs50009-100T","price":452.0,"currency_code":"USD","in_stock":true},{"title":"20T","offer_id":41683118522443,"sku":"abs50009-20T","price":151.0,"currency_code":"USD","in_stock":true}]},{"product_id":"mtt-cell-proliferation","title":"MTT Cell Proliferation and Cytotoxicity Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Place the cell plate in the sterile operating table.\u003cbr\u003e2. Add 10% volume of MTT solvent to each well of the cell plate. [Preparation of MTT solution: Dissolve 25mg of MTT with 5ml of MTT solvent to prepare a 5mg\/ml MTT solution. It can be used after preparation, or stored directly at-20 ℃ in the dark from light, or stored at-20 ℃ in the dark from light after appropriately packaging as needed.]\u003cbr\u003e3. Put the cell plate back into the incubator and continue to incubate for 3-4 hours. When doing comparative experiments, the incubation time should be consistent.\u003cbr\u003e4. After incubation, take out the cell plate from the incubator and dissolve it according to the following method to produce MTT formazan crystals.\u003cbr\u003e(1) If it is an adherent cell, discard the culture medium. Add the same amount of Formazan Solvent as the initial volume. The volume of dissolved fluid may vary per well, but the total volume must be consistent for comparison.\u003cbr\u003e(2) If the removal of non-adherent cells or culture medium will cause the loss of MTT formazan, add the same amount of lysate as the initial volume directly to the culture medium.\u003cbr\u003e(3) Readings must be completed within 1 hour after addition of Formazan Solvent.\u003cbr\u003e5. Gentle shaking in a shaker can enhance dissolution. In some cases, particularly when the cell density is high, repeated pipetting is required to completely dissolve the MTT formazan crystals.\u003cbr\u003e6. Spectrophotometer to measure OD570 and reference wavelength OD690.\u003cbr\u003e7. The 96-well plate can be detected on a microplate reader equipped with a suitable filter.\u003cbr\u003e8. Other types of multi-well plates need to be spectrophotometrically detected with a cuvette or plate reader of appropriate size.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eMTT Cell Proliferation and Cytotoxicity Assay Kit,MTT kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eThe MTT assay, also known as MTT colorimetry, is a method to detect cell survival and growth. The detection principle is that succinate dehydrogenase in mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-violet crystalline Formazan and deposit it in cells, while dead cells do not have this function. Within a certain range of cell numbers, the amount of MTT crystals formed is directly proportional to the number of cells. The greater the absorbance, the stronger the cell activity, indicating that the drug toxicity is less. This method has been widely used in the activity detection of some bioactive factors, large-scale anti-tumor drug screening, cytotoxicity test and tumor radiosensitivity determination. It is characterized by high sensitivity and economy.\u003c\/p\u003e\n\u003cdiv\u003eComponents: MTT (25mg), MTT solvent (5ml), Formazan solution (55ml)\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Since a 96-well plate is used for detection, if the cell culture time is long, we must pay attention to the problem of evaporation. On the one hand, since the circle around the 96-well plate is the easiest to evaporate, you can discard the circle around the plate and add PBS, water or culture medium instead; On the other hand, a 96-well plate can be placed close to a water source in the incubator to alleviate evaporation.\u003cbr\u003e2. If the test compound used can react with MTT, centrifuge first and then discard the culture medium. Carefully flush it with PBS for 2 to 3 times, and then add the culture medium containing MTT.\u003cbr\u003e3. The production of Formazan is not only proportional to the number of living cells, but also affected by the action time. Therefore, when there are many samples, the measurement results may change with time.\u003cbr\u003e4. This product contains carcinogens, avoid contact with skin and eyes, please operate carefully. For your safety and health, please operate in a lab coat and disposable gloves.\u003cbr\u003e5. This product is only used for scientific research and cannot be used clinically or diagnostically.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStored at 2 ~ 8 ℃, protected from light, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"500T","offer_id":41518128595019,"sku":"abs50010-500T","price":100.0,"currency_code":"USD","in_stock":true}]},{"product_id":"cell-proliferation-assay-kit","title":"MTS Cell Proliferation Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003eExperimental operation:\u003c\/strong\u003e\u003cbr\u003e\u003cstrong\u003e1. MTS working fluid preparation:\u003c\/strong\u003e\u003cbr\u003eMix MTS Solution and PMS Solution at a ratio of 20: 1 to obtain the MTS working solution. If the prepared MTS working solution needs to be stored, it should be stored at-20 ℃ in the dark from light. At this time, it is recommended to dispense appropriate amounts to avoid repeated freezing and thawing.\u003cbr\u003eNote: Both MTS Solution and PMS Solution are easy to decompose when exposed to light, so they need to be protected from light during preparation and use (can be wrapped in tin foil). If you want to store this kit at 4 °C for frequent use, the MTS working solution should be prepared before each test. The prepared unused MTS working solution needs to be stored at-20 °C protected from light.\u003cbr\u003e\u003cstrong\u003e2. Cell culture:\u003c\/strong\u003e\u003cbr\u003eThe cells are seeded in a suitable container such as a well plate. For 96-well plates, it is recommended to add 100uL of medium per well, and it is recommended to inoculate 5-100 × 10 per well\u003csup\u003e3\u003c\/sup\u003eCells. The cells of the experimental group can be treated with the drug of interest for a period of time according to the experimental design, at which time it is necessary to set up a blank group (containing medium only) and a control group (containing medium, cells and drug solvent).\u003cbr\u003eNote: The specific number of cells seeded per well can be adjusted according to the specific cell type.\u003cbr\u003e\u003cstrong\u003e3. MTS incubation:\u003c\/strong\u003e\u003cbr\u003e20 uL of MTS working solution was added to each well and then incubated in a cell incubator for 1-4 h.\u003cbr\u003eNote: The MTS incubation time can be adjusted for specific experiments.\u003cbr\u003e\u003cstrong\u003e4. Detection:\u003c\/strong\u003e\u003cbr\u003eThe absorbance (A) was measured at 490 nm with a microplate reader. If a 490 nm filter is not available, optionally use a 450-540 nm filter.\u003cbr\u003e\u003cstrong\u003e5. Data analysis:\u003cbr\u003e\u003c\/strong\u003eThe cell survival rate is calculated as follows\u003cbr\u003eCell viability (%) = [(A\u003csub\u003eExperimental group\u003c\/sub\u003e-A\u003csub\u003eBlank group\u003c\/sub\u003e)\/(A\u003csub\u003eControl group\u003c\/sub\u003e-A\u003csub\u003eBlank group\u003c\/sub\u003e)] ×100\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eMTS Cell Proliferation Assay Kit (MTS Cell Proliferation Assay Kit) is a common kit for detecting cell proliferation and toxicity. MTS is a novel tetrazolium salt, which can be reduced by succinate dehydrogenase in mitochondria of living cells to the water-soluble product Formazan. Formazan is specifically absorbed at 490 nm, so cell proliferation can be reflected by measuring the absorbance at 490 nm. The faster the cell proliferation, the higher the absorbance value.\u003cbr\u003eCompared with the common MTT kit, this kit does not need to remove the original culture medium, nor does it need to use another solvent to dissolve formazan, which is more convenient to use, and can avoid experimental errors caused by taking away part of formazan when removing the culture medium. This kit is very flexible to use, and incubation can be continued after detection to obtain better color development results. In addition, the kit has the advantages of wide linear range, high sensitivity and safe use.\u003cbr\u003eProduct Components:\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 52.5641%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.1426%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 12.4759%; text-align: center;\"\u003e500T\u003c\/td\u003e\n\u003ctd style=\"width: 10.6838%; text-align: center;\"\u003e2500T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.1426%; text-align: center;\"\u003eMTS Solution\u003c\/td\u003e\n\u003ctd style=\"width: 12.4759%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 10.6838%; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 24.1426%; text-align: center;\"\u003ePMS Solution\u003c\/td\u003e\n\u003ctd style=\"width: 12.4759%; text-align: center;\"\u003e0.5 mL\u003c\/td\u003e\n\u003ctd style=\"width: 10.6838%; text-align: center;\"\u003e2.5 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. When the cells are cultured for a long time, the edge wells of the 96-well plate are prone to liquid evaporation. It is recommended to discard the circle around the 96-well plate and add sterile water, culture medium or PBS instead; At the same time, the 96-well plate can be placed in the incubator close to the water source.\u003cbr\u003e2. MTS Solution and PMS Solutin will solidify at low temperatures. Before use, please return to room temperature or 37 °C water bath for about 10 minutes until completely melted before use.\u003cbr\u003e3. MTS Solution and PMS Solution are sensitive to light. If stored at 4 °C, the color of the solution may change slightly. This change may increase the background absorbance value, but it will not affect the final experimental results.\u003cbr\u003e4. If it cannot be detected in time, you can add 25uL of 10% SDS to each well after the incubation in Step 3 to terminate the reaction. The SDS-treated well plate can be placed\u003cbr\u003eStore in an incubator protected from light for no more than 18 hours.\u003cbr\u003e5. For your safety and health, please wear a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore the kit at -20°C away from light and moisture, stable for 1 year. For frequent use, the kit can be stored at 4°C away from light and moisture, stable for 6 weeks.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"1000T","offer_id":41518128627787,"sku":"abs50011-1000T","price":357.0,"currency_code":"USD","in_stock":true},{"title":"500T","offer_id":41683113115723,"sku":"abs50011-500T","price":164.0,"currency_code":"USD","in_stock":true},{"title":"2500T","offer_id":41683113148491,"sku":"abs50011-2500T","price":598.0,"currency_code":"USD","in_stock":true}]},{"product_id":"mitochondrial-membrane-potential-detection-kit","title":"Mitochondrial membrane potential detection kit (JC-1)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1 、 JC-1 Preparation of dyeing working solution \u003cbr\u003e Six-well plate required per well JC-1 The amount of dyeing working solution is 1mL , of other culture vessels JC-1 The amount of staining working solution used is similar: for the cell suspension per 50 ～ 100 Ten thousand cell needs 0.5mLJC-1 Dyeing Working Fluid. Take appropriate amount JC-1 （ 200× ), according to each 50μLJC-1 （ 200× ) join 8mL Proportional dilution of ultrapure water JC-1 。 Violently shake to fully dissolve and mix well JC-1 。 Then add 2mLJC-1 Staining buffer ( 5× ), after mixing, it is JC-1 Dyeing Working Fluid. \u003c\/p\u003e\n\u003cp\u003e2 Settings of Positive Control: \u003cbr\u003e Put the kit provided CCCP （ 10mM ) Recommended as per 1∶1000 The proportion is added to the cell culture solution, and diluted to 10μM , treating cells 20 Minutes. Subsequently loading as follows JC-1 , the mitochondrial membrane potential was detected. For most cells, typically 10μMCCCP dispose 20 After minutes, the membrane potential of mitochondria will be completely lost, JC-1 After staining, it should show green fluorescence; And normal cells through JC-1 Red fluorescence should be shown after staining. For specific cells, CCCP The concentration and duration of action may vary, so you need to refer to relevant literature to decide. \u003c\/p\u003e\n\u003cp\u003e3 For suspension cells, \u003cbr\u003e Take 10 ～ 60 Thousand cells, resuspended in 0.5mL In the cell culture medium, the cell culture medium may contain serum and phenol red. \u003cbr\u003e join 0.5mLJC-1 Dye the working solution, reverse it several times and mix well. In cell incubator 37℃ incubation 20 Minutes. \u003cbr\u003e During the incubation, according to each 1mLJC-1 Staining buffer ( 5× ) join 4mL Proportion of distilled water, prepare an appropriate amount JC-1 Staining buffer ( 1× ) and placed in an ice bath. \u003cbr\u003e37℃ After the incubation is completed, 600g4℃ Centrifugation 3 ～ 4 Min, the cells were pelleted. Discard the supernatant and be careful not to aspirate the cells as much as possible. \u003cbr\u003e use JC-1 Staining buffer ( 1× ) washing 2 Time: Join 1mLJC-1 Staining buffer ( 1× ) resuspending cells, 600g4℃ Centrifugation 3 ～ 4 Minutes, the cells were pelleted and the supernatant was discarded. \u003cbr\u003e Re-join 1mLJC-1 Staining buffer ( 1× ) resuspending cells, 600g4℃ Centrifugation 3 ～ 4 Minutes, the cells were pelleted and the supernatant was discarded. Use an appropriate amount JC-1 Staining buffer ( 1× ) After resuspension, observe with a fluorescence microscope or laser confocal microscope, or detect with a fluorescence spectrophotometer or analyze with a flow cytometer. \u003c\/p\u003e\n\u003cp\u003e4 For adherent cells, \u003cbr\u003e Note: For adherent cells, if you want to use fluorescence spectrophotometer or flow cytometer detection, you can collect the cells first, and refer to the detection method of suspended cells after resuspension. \u003cbr\u003e For one well of a six-well plate, the culture medium is aspirated and, if necessary, can be used according to the specific experiment PBS Or other suitable solution wash that cells once and add 1mL Cell culture fluid. The cell culture medium may contain serum and phenol red. join 1mLJC-1 Dyeing working solution, mix thoroughly. In cell incubator 37℃ incubation 20 Minutes. \u003cbr\u003e During the incubation, according to each 1mLJC-1 Staining buffer ( 5× ) join 4mL Proportion of distilled water, prepare an appropriate amount JC-1 Staining buffer ( 1× ) and placed in an ice bath. \u003cbr\u003e37℃ After the incubation, the supernatant was aspirated and treated with JC-1 Staining buffer ( 1× ) washing 2 Times. \u003cbr\u003e join 2mL The cell culture medium may contain serum and phenol red. \u003cbr\u003e Observe under fluorescence microscope or laser confocal microscope. \u003c\/p\u003e\n\u003cp\u003e5 For purified mitochondria \u003cbr\u003e Put the prepared JC-1 Reuse of dyeing working solution JC-1 Staining buffer ( 1× ) dilution 5 Times. \u003cbr\u003e0.9mL5 Double diluted JC-1 Add to the dyeing working solution 0.1mL The total protein content was 10 ～ 100μg Purified mitochondria. Detection with a fluorescence spectrophotometer or a fluorescence microplate reader: After mixing, time scan with a fluorescence spectrophotometer directly ( timescan ), the excitation wavelength is 485nm , emission wavelength is 590nm 。 If a fluorescent microplate reader is used, the excitation wavelength cannot be set to 485nm When, you can 475 ～ 520nm The excitation wavelength is set within the range. In addition, you can also refer to the following steps 6 The wavelength setting in is performed for fluorescence detection. \u003cbr\u003e Observation by fluorescence microscope or laser confocal microscope: the method is the same as the following steps 6 。 \u003c\/p\u003e\n\u003cp\u003e6 , fluorescence observation and result analysis detection \u003cbr\u003eJC-1 The excitation light can be set to 490nm , the emitted light is set to 530nm ； Detection JC-1 Polymer, the excitation light may be set to 525nm , the emitted light is set to 590nm 。 Note that it is not necessary to set the excitation light and the emission light at the maximum excitation wavelength and the maximum emission wavelength when measuring fluorescence here. If observed using a fluorescence microscope, detect JC-1 When monomer, you can refer to the settings when observing other green fluorescence, such as observing GFP Or FITC Time setting; Detection JC-1 Polymer may be observed with reference to other red fluorescence such as propidium iodide or Cy3 When setting. The appearance of green fluorescence indicates that the mitochondrial membrane potential decreases, and the cell is probably in the early stage of apoptosis. The appearance of red fluorescence indicates that the mitochondrial membrane potential is relatively normal and the state of cells is also relatively normal. \u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cdiv style=\"width: 100%; white-space: normal;\"\u003e    Mitochondrial membrane potential detection kit ( JC-1 ) is a method with JC-1 It is a fluorescent probe, which can quickly and sensitively detect the changes of membrane potential of cells, tissues or purified mitochondria, and can be used for early detection of apoptosis. \u003cbr\u003e   JC-1 Is a widely used method for detecting mitochondrial membrane potential △Ψm Ideal fluorescent probe. Cellular, tissue, or purified mitochondrial membrane potentials can be detected. When mitochondrial membrane potential is high, JC-1 Aggregate in the matrix of mitochondria, forming polymers that can   Producing red fluorescence; When mitochondrial membrane potential is low, JC-1 Cannot aggregate in the matrix of mitochondria, at which time JC-1 It is a monomer and can produce green fluorescence. In this way, it is very convenient to detect the change of mitochondrial membrane potential through the change of fluorescence color. The relative ratio of red and green fluorescence is commonly used to measure the ratio of mitochondrial depolarization.  \u003cbr\u003e    The decrease of mitochondrial membrane potential is a landmark event in the early stage of apoptosis. Pass JC-1 From red fluorescence   The transition to green fluorescence can easily detect the decrease in cell membrane potential, and can also be detected with JC-1 From red fluorescence to green   The transformation of color fluorescence is used as a detection index of early apoptosis.  \u003cbr\u003e   JC-1 The maximum excitation wavelength of the monomer is 515nm , the maximum emission wavelength is 529nm ； JC-1 The maximum of polymer   The excitation wavelength is 585nm , the maximum emission wavelength is 590nm 。 In actual observation, the conventional observation of red fluorescence and green fluorescence is used   Just setting it. \u003cbr\u003e    This kit provides CCCP As a positive control to induce a decrease in mitochondrial membrane potential.  \u003cbr\u003e    For samples in six-well plates, this kit can detect a total 100 Samples; For 12 The sample in the well, this kit can   To detect 200 Samples. \u003c\/div\u003e\n\u003cdiv\u003e\n\u003cp class=\"15\" style=\"margin-bottom: 7.8000pt; margin-left: 21.0000pt; mso-para-margin-bottom: 0.5000gd; text-indent: -21.0000pt; line-height: 20.0000pt; mso-line-height-rule: exactly; mso-list: l0 level1 lfo1;\"\u003e Product composition :\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 65.3595%; height: 159px; margin: 0 auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49%; text-align: center;\"\u003e\u003cstrong\u003e Product Name \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 50.8%; text-align: center;\"\u003e\u003cp\u003e\u003cstrong\u003e package \u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49%;\"\u003e\u003cp\u003eJC-1(200×)\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 50.8%;\"\u003e\u003cp\u003e100ul\/ Tube, total  5  Tube \u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49%;\"\u003e\u003cp\u003e Ultrapure water \u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 50.8%;\"\u003e\u003cp\u003e90mL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49%;\"\u003e\u003cp\u003eJC-1  Staining buffer   (5×)\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 50.8%;\"\u003e\u003cp\u003e80mL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49%;\"\u003e\u003cp\u003eCCCP(10mM)\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 50.8%;\"\u003e\u003cp\u003e20ul\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cdiv align=\"center\"\u003e \u003c\/div\u003e\n\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. JC-1 (200 ×) will solidify and stick to the bottom, wall or cover of the centrifuge tube at lower temperatures such as 4 °C and ice bath. It can be incubated in a water bath at 20 ~ 25 °C for a while until it is completely melted. Use later.\u003cbr\u003e2. JC-1 (200 ×) must be fully dissolved and mixed with the ultrapure water provided by the kit before JC-1 staining buffer (5 ×) can be added. Do not prepare JC-1 staining buffer (1 ×) first and then add JC-1 (200 ×), so JC-1 will be difficult to fully dissolve, which will seriously affect subsequent detection.\u003cbr\u003e3. When washing with JC-1 staining buffer (1 ×) after loading JC-1, keep the JC-1 staining buffer (1 ×) at about 4 ℃, and the washing effect at this time is better.\u003cbr\u003e4. After the JC-1 probe is loaded and washed, try to complete the follow-up detection within 30 minutes. Store in ice bath before testing.\u003cbr\u003e5. Do not prepare all JC-1 staining buffer (5 ×) into JC-1 staining buffer (1 ×). JC-1 staining buffer (5 ×) should be directly used during the use of this kit.\u003cbr\u003e6. If precipitate is found in JC-1 staining buffer (5 ×), it must be completely dissolved before it can be used. To promote dissolution, it can be heated at 37 °C.\u003cbr\u003e7. CCCP is a mitochondrial electron transport chain inhibitor, which is toxic. Please pay attention to careful protection.\u003cbr\u003e8. For your safety and health, please wear a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eStore at 4 ℃, protected from light, shelf life for one year\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100T","offer_id":41518130069579,"sku":"abs50016-100T","price":150.0,"currency_code":"USD","in_stock":true}]},{"product_id":"mitochondrial-membrane-potential-assay-kit","title":"Mitochondrial Membrane Potential Detection Kit (jc-10)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Preparation of JC-10 dyeing working solution:\u003cbr\u003eThe amount of JC-10 staining working solution required for each well of the six-well plate is 1 mL, and the amount of JC-10 staining working solution for other culture vessels is similar; For the cell suspension, 0.5 mL of JC-10 staining working solution is required for every 500,000 to 1,000,000 cells. Take an appropriate amount of JC-10 (200 ×) and dilute JC-10 in the proportion of 8mL of ultrapure water per 50μL of JC-10 (200 ×). Thoroughly dissolve and mix JC-10 with vigorous shaking. Then add 2mL of JC-10 staining buffer (5 ×), and mix well to obtain JC-10 staining working solution.\u003cbr\u003e2. Setting of positive control:\u003cbr\u003eCCCP (10 mM) provided in the kit is recommended to be added to the cell culture medium at a ratio of 1: 1000, diluted to 10 μM, and treated the cells for 20 minutes. Subsequently, JC-10 was loaded according to the following method, and mitochondrial membrane potential was detected. For most cells, the membrane potential of mitochondria will usually be completely lost after 20 minutes of 10 μM CCCP treatment, and it should show green fluorescence after JC-10 staining; However, normal cells should show red fluorescence after staining with JC-10. For specific cells, the concentration and duration of CCCP may be different, and it is necessary to refer to relevant literature to decide.\u003cbr\u003e3. For suspension cells:\u003cbr\u003e(1) Take 100,000 to 600,000 cells and resuspend them in 0.5 mL cell culture medium. The cell culture medium can contain serum and phenol red.\u003cbr\u003e(2) Add 0.5 mL of JC-10 dyeing working solution, reverse it several times and mix well. Incubate in a cell incubator at 37 °C for 20 minutes.\u003cbr\u003e(3) During the incubation period, an appropriate amount of JC-10 staining buffer (1 ×) was prepared according to the ratio of adding 4 mL of distilled water per 1 mL of JC-10 staining buffer (5 ×), and placed in an ice bath.\u003cbr\u003e(4) After the incubation at 37 °C, 600g was centrifuged at 4 °C for 3-4 minutes to precipitate the cells. Discard the supernatant and be careful not to aspirate the cells as much as possible.\u003cbr\u003e(5) Wash twice with JC-10 staining buffer (1 ×): add 1 mL of JC-10 staining buffer (1 ×) to resuspend the cells, centrifuge 600 g at 4 °C for 3-4 minutes, pellet the cells, and discard the supernatant. Add 1 mL of JC-10 staining buffer (1 ×) to resuspend the cells, centrifuge 600g at 4 °C for 3-4 minutes, pellet the cells, and discard the supernatant.\u003cbr\u003e(6) After resuspending with an appropriate amount of JC-10 staining buffer (1 ×), observe with a fluorescence microscope or laser confocal microscope, or detect with a fluorescence spectrophotometer or analyze with a flow cytometer.\u003cbr\u003e4. For adherent cells:\u003cbr\u003eNote: For adherent cells, if you want to use fluorescence spectrophotometer or flow cytometer detection, you can collect the cells first, and refer to the detection method of suspended cells after resuspension.\u003cbr\u003e(1) For one well of a six-well plate, aspirate the culture medium, wash the cells once with PBS or other appropriate solution if necessary according to the specific experiment, and add 1 mL of cell culture medium. The cell culture medium may contain serum and phenol red.\u003cbr\u003e(2) Add 1mL of JC-10 dyeing working solution and mix well. Incubate in a cell incubator at 37 °C for 20 minutes.\u003cbr\u003e(3) During the incubation period, an appropriate amount of JC-10 staining buffer (1 ×) was prepared according to the ratio of adding 4 mL of distilled water per 1 mL of JC-10 staining buffer (5 ×), and placed in an ice bath.\u003cbr\u003e(4) After the incubation at 37 °C, the supernatant was aspirated and washed twice with JC-10 staining buffer (1 ×).\u003cbr\u003e(5) 2 mL of cell culture medium is added, and the culture medium may contain serum and phenol red.\u003cbr\u003e(6) Observe under fluorescence microscope or laser confocal microscope.\u003cbr\u003e5. For purified mitochondria:\u003cbr\u003e(1) The prepared JC-10 staining working solution was diluted 5 times with JC-10 staining buffer (1 ×).\u003cbr\u003e(2) 0.9 mL of 5-fold diluted JC-10 staining working solution was added with 0.1 mL of purified mitochondria with a total protein amount of 10-100 μg.\u003cbr\u003e(3) Detection with a fluorescence spectrophotometer or a fluorescence microplate reader: After mixing, directly use a fluorescence spectrophotometer to time scan (time scan). The excitation wavelength is 485nm and the emission wavelength is 590nm. If a fluorescent microplate reader is used and the excitation wavelength cannot be set to 485 nm, the excitation wavelength can be set in the range of 475 to 520 nm. In addition, fluorescence detection can also be performed with reference to the wavelength setting in step 6 below.\u003cbr\u003e(4) Observation with fluorescence microscope or laser confocal microscope: The method is the same as step 6 below.\u003cbr\u003e6. Fluorescence observation and result analysis:\u003cbr\u003eWhen detecting JC-10 monomer, the excitation light can be set to 490nm and the emission light can be set to 530nm; When detecting the JC-10 polymer, the excitation light can be set to 525 nm and the emission light can be set to 590 nm. Note that it is not necessary to set the excitation light and the emission light at the maximum excitation wavelength and the maximum emission wavelength when measuring fluorescence here. For observation using a fluorescence microscope, when detecting JC-10 monomer, you can refer to the settings when observing other green fluorescence, such as the settings when observing GFP or FITC; When detecting JC-10 polymer, you can refer to the settings when observing other red fluorescence, such as propidium iodide or Cy3. The appearance of green fluorescence indicates that the mitochondrial membrane potential decreases, and the cell is probably in the early stage of apoptosis. The appearance of red fluorescence indicates that the mitochondrial membrane potential is relatively normal and the state of cells is also relatively normal.\u003cbr\u003e\u003cbr\u003e\u003cbr\u003ePrecautions:\u003cbr\u003e1. JC-10 (200 ×) will solidify and stick to the bottom, wall or cover of the centrifuge tube at lower temperatures such as 4 °C and ice bath. It can be incubated in a water bath at 20 ~ 25 °C for a while until it is completely melted. After use.\u003cbr\u003e2. JC-10 (200 ×) must be fully dissolved and mixed with the ultrapure water provided by the kit before JC-10 staining buffer (5 ×) can be added. Do not prepare JC-10 staining buffer (1 ×) first and then add JC-10 (200 ×), as it will be difficult to fully dissolve JC-10, which will seriously affect subsequent detection.\u003cbr\u003e3. When washing with JC-10 staining buffer (1 ×) after loading JC-10, keep the JC-10 staining buffer (1 ×) at about 4 ℃, and the washing effect at this time is better.\u003cbr\u003e4. After the JC-10 probe is loaded and washed, try to complete the follow-up detection within 30 minutes. Store in ice bath before testing.\u003cbr\u003e5. Please do not prepare all JC-10 staining buffer (5 ×) into JC-10 staining buffer (1 ×). JC-10 staining buffer (5 ×) should be directly used during the use of this kit.\u003cbr\u003e6. If precipitate is found in JC-10 staining buffer (5 ×), it must be completely dissolved before use. To promote dissolution, it can be heated at 37 °C.\u003cbr\u003e7. CCCP is a mitochondrial electron transport chain inhibitor, which is toxic. Please pay attention to careful protection.\u003cbr\u003e8. For your safety and health, please wear a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eMitochondrial membrane potential detection kit (JC-10) is a kit that uses JC-10 as a fluorescent probe to rapidly and sensitively detect the changes of mitochondrial membrane potential of cells, tissues or purified mitochondria. It can be used for early apoptosis detection.\u003cbr\u003eJC-10 is an ideal fluorescent probe widely used to detect mitochondrial membrane potential ΔΨm. Cellular, tissue, or purified mitochondrial membrane potentials can be detected. When the mitochondrial membrane potential is high, JC-10 gathers in the matrix of mitochondria and forms a polymer, which can produce red fluorescence; When the mitochondrial membrane potential is low, JC-10 cannot gather in the mitochondrial matrix. At this time, JC-10 is a monomer and can produce green fluorescence. In this way, it is very convenient to detect the change of mitochondrial membrane potential through the change of fluorescence color. The relative ratio of red and green fluorescence is commonly used to measure the ratio of mitochondrial depolarization.\u003cbr\u003eThe decrease of mitochondrial membrane potential is a landmark event in the early stage of apoptosis. The decrease of cell membrane potential can be easily detected by the transition from red fluorescence to green fluorescence of JC-10, and the transition from red fluorescence to green fluorescence of JC-10 can also be used as a detection index in the early stage of apoptosis.\u003cbr\u003eThe maximum excitation wavelength of JC-10 monomer is 515nm, and the maximum emission wavelength is 529nm; The JC-10 polymer has a maximum excitation wavelength of 585 nm and a maximum emission wavelength of 590 nm. In actual observation, conventional settings for observing red fluorescence and green fluorescence may be used.\u003cbr\u003eThis kit provides CCCP as a positive control for inducing a decrease in mitochondrial membrane potential. For samples in six-well plates, this kit can detect a total of 100 samples; For samples in 12 wells, this kit can detect a total of 200 samples.\u003cbr\u003e\u003cbr\u003eProduct Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 33.4482%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.8408%;\"\u003eComponent Name\u003c\/td\u003e\n\u003ctd style=\"width: 11.6766%;\"\u003eSpecifications\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.8408%;\"\u003eJC-10 (200 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 11.6766%;\"\u003e100 μL\/tube, 5 tubes in total\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.8408%;\"\u003eUltrapure water\u003c\/td\u003e\n\u003ctd style=\"width: 11.6766%;\"\u003e90mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.8408%;\"\u003eJC-10 Staining Buffer (5 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 11.6766%;\"\u003e80mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 10.8408%;\"\u003eCCCP (10 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 11.6766%;\"\u003e20 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eDry and protected from light at-20 ℃, shelf life is 12 months. Ultrapure water and JC-10 staining buffer (5 ×) can also be stored at 4 °C.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100T","offer_id":41518130102347,"sku":"abs50017-100T","price":450.0,"currency_code":"USD","in_stock":true}]},{"product_id":"caspase-1","title":"Caspase 1 Activity Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Preparatory work\u003cbr\u003e1. After the lysate is dissolved, mix well and put on an ice bath for later use.\u003cbr\u003e2. After the detection buffer is dissolved, mix well and place it on an ice bath for later use.\u003cbr\u003e\u003cbr\u003e2. Determination of pNA standard curve\u003cbr\u003e1. Preparation of standard diluent: Prepare an appropriate amount of standard diluent according to the ratio of 0.1 ml of lysate per 0.9 ml of detection buffer.\u003cbr\u003e2. The pNA provided in the kit (10 mM) was diluted to 0, 10, 20, 50, 100 and 200 uM with a standard diluent as a standard.\u003cbr\u003e3. Take 100ul of each concentration and detect it with a microplate reader, or take an appropriate amount and detect it with a spectrophotometric detection cup with a capacity of no more than 100ul to determine A405.\u003cdiv\u003e4. A405 of each standard was subtracted from A405 of the blank control without pNA to calculate the actual absorbance due to pNA, and a standard curve of pNA concentration versus A405 was created. The pNA standard curve has a good linear relationship in the range of 0-20uM, as shown in the figure below.\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240329\/3fd22d866bc94b129b0a094532b79d84.png\" alt=\"\" width=\"438\" height=\"324\"\u003eNote: The experimental data may vary due to different experimental conditions, testing instruments, etc. The data in the figure is for reference only.\u003cbr\u003e\u003cbr\u003e3. Sample collection\u003cbr\u003e1. Suspended cells: Centrifuge the control sample that did not induce apoptosis and the sample that induced apoptosis at 600g at 4 º C for 5 minutes to collect the cells, carefully aspirate the supernatant, and ensure that no cells are aspirated as much as possible, and wash once with PBS. After the supernatant was aspirated as before, the lysate was added at a ratio of 100 ul of lysate per 2 million cells, the pellet was resuspended, and lysed in an ice bath for 15 minutes. Go down to Step 4.\u003cbr\u003e2. Adherent cells: suck the cell culture medium for later use. The adherent cells were digested with trypsin and collected into a spare cell culture medium. Collect cells by centrifugation at 600 g at 4 º C for 5 minutes, carefully aspirate the supernatant while ensuring that no cells are aspirated as much as possible, and wash once with PBS. After the supernatant was aspirated as before, the lysate was added at a ratio of 100 ul of lysate per 2 million cells, the pellet was resuspended, and lysed in an ice bath for 15 minutes. Go down to Step 4.\u003cbr\u003e3. Tissue samples: Add lysate at a ratio of 100ul lysate per 3-10mg tissue, and homogenize with a glass homogenizer on an ice bath. The homogenate was then transferred to a 1.5 ml centrifuge tube and lysed in an ice bath for an additional 5 minutes.\u003cbr\u003e4. Centrifuge at 4 º C 16,000-20,000 g for 10-15 minutes.\u003cbr\u003e5. Transfer the supernatant to an ice bath precooled centrifuge tube.\u003cbr\u003e6. The enzyme activity of Caspase 1 was determined immediately or the sample was stored at − 80 ° C. At the same time, a small amount of sample can be taken to measure the protein concentration by Bradford method, and try to make the protein concentration reach 1-3mg\/ml, which is equivalent to at least 10-30ug of protein per 10ul of the sample to be tested. If there are fewer cells, the amount of cells can be appropriately increased.\u003cbr\u003e\u003cbr\u003e4. Detection of Caspase 1 enzyme activity\u003cbr\u003e1. Take out an appropriate amount of Ac-YVAD-pNA (2mM) and place it on an ice bath for later use.\u003cbr\u003e2. Set up the reaction system as follows:\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 92.0261%; height: 125px; margin: 0px auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; text-align: center; height: 22px;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; text-align: center; height: 22px;\"\u003eBlank control\u003c\/td\u003e\n\u003ctd style=\"width: 31.491%; text-align: center; height: 22px;\"\u003eSample\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003eAssay buffer\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003e40ul\u003c\/td\u003e\n\u003ctd style=\"width: 31.491%; height: 22px;\"\u003e40ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003eSample to be measured\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003e0ul\u003c\/td\u003e\n\u003ctd style=\"width: 31.491%; height: 22px;\"\u003e50ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003eLysate\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003e50ul\u003c\/td\u003e\n\u003ctd style=\"width: 31.491%; height: 22px;\"\u003e0ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003eAc-YVAD-pNA (2 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 22px;\"\u003e10ul\u003c\/td\u003e\n\u003ctd style=\"width: 31.491%; height: 22px;\"\u003e10ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 15px;\"\u003e\n\u003ctd style=\"width: 31.4869%; height: 15px;\"\u003eTotal Volume\u003c\/td\u003e\n\u003ctd style=\"width: 31.4869%; height: 15px;\"\u003e100ul\u003c\/td\u003e\n\u003ctd style=\"width: 31.491%; height: 15px;\"\u003e100ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003eNote: When setting the reaction system, add the detection buffer first, then add the sample to be tested, and mix properly. Be careful to avoid bubbles during mixing. An additional 10 ul of Ac-YVAD-pNA (2 mM) was then added.\u003cbr\u003e\u003cbr\u003e3. Add Ac-YVAD-pNA (2mM) and mix well, taking care to avoid bubbles during mixing. Incubate at 37 º C for 60-120 minutes. A405 can be measured when the color change is obvious. If the color change is not obvious, the incubation time can be appropriately extended, or even overnight.\u003cbr\u003e4. A405 of the sample minus A405 of the blank control is the absorbance produced by pNA catalyzed by Caspase 1 in the sample. The amount of pNA catalytically produced in the sample can be calculated by comparing it with the standard curve obtained in step 2.\u003cbr\u003e5. Refer to the definition of Caspase 1 enzyme activity unit from Chemicon: One unit is the amount of enzyme that will clear 1.0 nmol of the colorimetric substrate Ac-YVAD-pNA per hour at 37 º C under saturated substrate concentrations. That is, an enzyme activity unit is defined as the amount of caspase 1 enzyme that can cleave 1 nmol of Ac-YVAD-pNA to produce 1 nmol of pNA within one hour at 37 º C when the substrate is saturated. In this way, it is possible to calculate how many enzyme activity units of Caspase 1 are contained in the sample. Note: In the detection system of this kit, the initial concentration of the substrate is 0.2 mM, at which time the substrate is saturated. For many samples, the substrate is saturated within 2 hours of incubation at 37 º C; If the activity of caspase 1 enzyme in the sample is particularly high, the sample must be properly diluted with lysate before determination.\u003cbr\u003e6. Use the Bradford method to detect the protein concentration in the sample to be tested (because the lysate contains a high concentration of DTT, it is not suitable to use the BCA method for protein concentration determination). In this way, the enzyme activity unit of Caspase 1 contained per unit weight of protein in a sample can be calculated.\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003einterkeukin 1β conberting enzyme,ICE,caspase-1,caspase 1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eMammalian tissues, cells\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eCaspase 1 Activity Detection Kit is a kit for detecting Caspase 1 enzyme activity or purified Caspase 1 enzyme activity in cell or tissue lysates by spectrophotometry. Caspase is a family of proteases involved in the process of apoptosis and contains more than 10 members. Among them, Caspase-1 is the only Caspase that can cleave IL-1b and IL-18 precursors to produce active cytokines. Caspase-11 cleaves the 45 kD Caspase-1 precursor protein to produce 20 kD and 10 kD fragments, which can form heterodimers and further tetramers. This tetramer has protease activity. Caspase-1 regulates apoptosis by cleaving Bcl-XL, and regulates related cytokine-mediated immune inflammatory responses by cleaving cytokine precursors.\u003cbr\u003eThis kit is based on the fact that Caspase 1 can catalyze the substrate Ac-YVAD-pNA to produce yellow pNA (p-nitroaniline), so that the activity of Caspase 1 can be detected by measuring absorbance. pNA has strong absorption near 405nm. This kit can detect 20 samples (20T) or 100 samples (100T) in addition to the standard curve when detected with a microplate reader or a spectrophotometric detection cup with a capacity not exceeding 100ul.\u003cbr\u003e\u003cstrong\u003eProduct Components:\u003cbr\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 62.7886%; margin: 0 auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.4969%; text-align: center;\"\u003eProduct composition\u003c\/td\u003e\n\u003ctd style=\"width: 14.453%; text-align: center;\"\u003e20T\u003c\/td\u003e\n\u003ctd style=\"width: 11.3903%; text-align: center;\"\u003e100T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.4969%; text-align: center;\"\u003eLysate Lysis buffer\u003c\/td\u003e\n\u003ctd style=\"width: 14.453%; text-align: center;\"\u003e8ml\u003c\/td\u003e\n\u003ctd style=\"width: 11.3903%; text-align: center;\"\u003e30ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.4969%; text-align: center;\"\u003eAssay buffer\u003c\/td\u003e\n\u003ctd style=\"width: 14.453%; text-align: center;\"\u003e8ml\u003c\/td\u003e\n\u003ctd style=\"width: 11.3903%; text-align: center;\"\u003e20ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.4969%; text-align: center;\"\u003eAc-YVAD-pNA (2 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 14.453%; text-align: center;\"\u003e200ul\u003c\/td\u003e\n\u003ctd style=\"width: 11.3903%; text-align: center;\"\u003e1ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.4969%; text-align: center;\"\u003epNA (10 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 14.453%; text-align: center;\"\u003e200ul\u003c\/td\u003e\n\u003ctd style=\"width: 11.3903%; text-align: center;\"\u003e1ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eWhen stored at-20 °C, Ac-YVAD-pNA and pNA need to be stored in the dark.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"20T","offer_id":41518130135115,"sku":"abs50023-20T","price":117.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41518130167883,"sku":"abs50023-100T","price":333.0,"currency_code":"USD","in_stock":true}]},{"product_id":"caspase-3-7","title":"Caspase 3 \/ 7 Activity Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eOperating Instructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eIn the process of using PBS, special care should be taken to avoid microbial contamination of the solution. PBS may produce precipitation at low temperatures. If precipitate is found, place in a 37 °C water bath until completely dissolved before use. For your safety and health, please operate in a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e\u003cspan style=\"font-size: 14pt;\"\u003eInstructions for Use:\u003c\/span\u003e\u003cbr\u003e1. Preparation:\u003c\/strong\u003e\u003cbr\u003eA. After the lysate is dissolved, mixed well and placed on an ice bath for later use.\u003cbr\u003eb. The test buffer is dissolved and mixed well and placed on an ice bath for later use.\u003cbr\u003e\u003cstrong\u003e2. Determination of pNA standard curve:\u003c\/strong\u003e\u003cbr\u003eA. Preparation of standard diluent: Prepare an appropriate amount of standard diluent according to the ratio of 0.1 ml of lysate per 0.9 ml of detection buffer.\u003cbr\u003eb. The pNA (10 mM) supplied in the kit was diluted to 0, 10, 20, 50, 100 and 200 μM with a standard diluent as a standard.\u003cbr\u003ec. Take 100μl of each concentration and detect it with a microplate reader, or take an appropriate amount and detect it with a spectrophotometric detection cup with a capacity not exceeding 100μl to determine A405.\u003cbr\u003ed. The actual absorbance due to pNA was calculated by subtracting the A405 of the blank control without pNA from the A405 of each standard, and a standard curve of pNA concentration versus A405 was created.\u003cbr\u003e\u003cstrong\u003e3. Collection of samples:\u003c\/strong\u003e\u003cbr\u003eA. For suspended cells: Centrifuge the control sample that did not induce apoptosis and the sample that induced apoptosis, 600 g at 4 º C for 5 minutes to collect the cells, carefully aspirate the supernatant, and ensure that no cells are aspirated as much as possible, and wash once with PBS. After the supernatant was aspirated as before, the lysate was added at a ratio of 100 microliters of lysate per 2 million cells, the pellet was resuspended, and lysed in an ice bath for 15 minutes. Go down to step 3d.\u003cbr\u003eb. For adherent cells: aspirate the cell culture medium for later use. The adherent cells were digested with trypsin and collected into a spare cell culture medium. Cells were collected by centrifugation at 600 g at 4 º C for 5 minutes, the supernatant was carefully aspirated, while ensuring that no cells were aspirated as much as possible, and PBS was washed once. After the supernatant was aspirated as before, the lysate was added at a ratio of 100 microliters of lysate per 2 million cells, the pellet was resuspended, and lysed in an ice bath for 15 minutes. Go down to step 3d.\u003cbr\u003ec. For tissue samples: Lysate was added at a ratio of 100 microliters of lysate per 3-10 mg of tissue and homogenized with a glass homogenizer on an ice bath. The homogenate was then transferred to a 1.5 ml centrifuge tube and lysed in an ice bath for an additional 5 minutes.\u003cbr\u003ed. Centrifuge at 4 º C 16,000-20,000 g for 10-15 minutes.\u003cbr\u003ee. The supernatant was transferred to an ice bath precooled centrifuge tube.\u003cbr\u003ef. Determine the enzyme activity of caspase 3\/7 immediately or save the sample at-80 º C. At the same time, a small amount of sample can be taken to measure the protein concentration by Bradford method, and try to make the protein concentration reach 1-3mg\/ml, which is equivalent to at least 10-30μg of protein per 10 microliters of sample to be tested. If there are fewer cells, the amount of cells can be appropriately increased.\u003cbr\u003e\u003cstrong\u003e4. Detection of Caspase 3\/7 enzyme activity:\u003c\/strong\u003e\u003cbr\u003eA. Remove an appropriate amount of Ac-DEVD-pNA (2 mM) and place it on an ice bath for later use.\u003cbr\u003eb. The reaction system was set up as follows:\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 60.9187%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 21.8354%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 15.7597%; text-align: center;\"\u003eBlank control\u003c\/td\u003e\n\u003ctd style=\"width: 15.9566%; text-align: center;\"\u003eSample\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 21.8354%; text-align: center;\"\u003eAssay buffer\u003c\/td\u003e\n\u003ctd style=\"width: 15.7597%; text-align: center;\"\u003e40 μl\u003c\/td\u003e\n\u003ctd style=\"width: 15.9566%; text-align: center;\"\u003e40 μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 21.8354%; text-align: center;\"\u003eSample to be measured\u003c\/td\u003e\n\u003ctd style=\"width: 15.7597%; text-align: center;\"\u003e0 μl\u003c\/td\u003e\n\u003ctd style=\"width: 15.9566%; text-align: center;\"\u003e50 μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 21.8354%; text-align: center;\"\u003eLysate\u003c\/td\u003e\n\u003ctd style=\"width: 15.7597%; text-align: center;\"\u003e50 μl\u003c\/td\u003e\n\u003ctd style=\"width: 15.9566%; text-align: center;\"\u003e0 μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 21.8354%; text-align: center;\"\u003eAc-DEVD-pNA (2 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 15.7597%; text-align: center;\"\u003e10 μl\u003c\/td\u003e\n\u003ctd style=\"width: 15.9566%; text-align: center;\"\u003e10 μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 21.8354%; text-align: center;\"\u003eTotal Volume\u003c\/td\u003e\n\u003ctd style=\"width: 15.7597%; text-align: center;\"\u003e100 μl\u003c\/td\u003e\n\u003ctd style=\"width: 15.9566%; text-align: center;\"\u003e100 μl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003eNote: When setting the reaction system, add the detection buffer first, then add the sample to be tested, and mix properly. Be careful to avoid bubbles during mixing. An additional 10 μl of Ac-DEVD-pNA (2 mM) was then added.\u003cstrong\u003e\u003cbr\u003e\u003cbr\u003e\u003c\/strong\u003ec. Add Ac-DEVD-pNA (2 mM) and mix well, taking care to avoid bubbles during mixing. Incubate at 37 º C for 60-120 minutes. A405 can be measured when the color change is obvious. If the color change is not obvious, the incubation time can be appropriately extended, or even overnight.\u003cbr\u003ed. A405 of the sample minus A405 of the blank control is the absorbance generated by pNA catalyzed by caspase 3\/7 in the sample. The amount of pNA catalytically produced in the sample can be calculated by comparing it with the standard curve obtained in step 1.\u003cbr\u003ee. Refer to the definition of caspase 3\/7 enzyme activity units from Chemicon: One unit is the amount of enzyme that will clear 1.0 nmol of the colorimetric substrate Ac-DEVD-pNA per hour at 37 º C under saturated substrate concentrations. That is, an enzyme activity unit is defined as the amount of caspase 3\/7 enzyme that can cleave 1 nmol of Ac-DEVD-pNA to produce 1 nmol of pNA within one hour at 37 º C when the substrate is saturated. This allows to calculate how many enzyme activity units of caspase 3\/7 are contained in the sample. Note: In the detection system of this kit, the initial concentration of the substrate is 0.2 mM, at which time the substrate is saturated. For many samples, the substrate is saturated within 2 hours of incubation at 37 º C; If the activity of caspase 3\/7 enzyme in the sample is particularly high, the sample must be properly diluted with lysate before determination.\u003cbr\u003ef. Use the Bradford method to detect the protein concentration in the sample to be tested (because the lysate contains a high concentration of DTT, it is not suitable to use the BCA method for protein concentration determination). In this way, the enzyme activity unit of caspase 3\/7 contained per unit weight of protein in a sample can be calculated.\u003cstrong\u003e\u003cbr\u003e\u003cbr\u003e\u003c\/strong\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCPP32,Yama,apopain,caspase-3,caspase 3,Activated caspase-3\/7,Cleaved caspase-3\/7,caspase-3\/7\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp style=\"font-weight: 400;\"\u003ecaspase 3\/7 Activity Assay Kit (caspase 3\/7 Activity Assay Kit) is a kit for detecting caspase 3\/7 enzyme activity or purified caspase 3\/7 enzyme activity in cell or tissue lysates by spectrophotometry.\u003cbr\u003eCaspase (Cysteine-requiring Aspartate Protease) is a family of proteases that play an important role in the process of apoptosis. caspase 3, also known as CPP32, Yama or apopain, sometimes written as caspase-3 or caspase 3, belongs to the CED-3 subfamily of the caspase family and is a key enzyme in the process of apoptosis. caspase 3 is the most studied caspase in mammalian cells. Both Caspase 3 and Caspase 7 can cleave the DEVD peptide sequence, so this kit can detect caspase3\/7.\u003cbr\u003eThis kit is based on the fact that caspase 3\/7 can catalyze the substrate Ac-DEVD-pNA to produce yellow pNA (p-nitroaniline), so that the activity of caspase 3\/7 can be detected by measuring absorbance. pNA has strong absorption near 405nm.\u003cbr\u003eThis kit can detect 20 samples (20T) or 100 samples (100T) in addition to the standard curve when detected with a microplate reader or a spectrophotometric detection cup with a capacity not exceeding 100μl.\u003cbr\u003e\u003cbr\u003eProduct Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 58.4211%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 11.5689%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 8.49085%; text-align: center;\"\u003e20T\u003c\/td\u003e\n\u003ctd style=\"width: 8.45254%; text-align: center;\"\u003e100T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 11.5689%; text-align: center;\"\u003eLysate Lysis buffer\u003c\/td\u003e\n\u003ctd style=\"width: 8.49085%; text-align: center;\"\u003e8ml\u003c\/td\u003e\n\u003ctd style=\"width: 8.45254%; text-align: center;\"\u003e30ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 11.5689%; text-align: center;\"\u003eAssay buffer\u003c\/td\u003e\n\u003ctd style=\"width: 8.49085%; text-align: center;\"\u003e8ml\u003c\/td\u003e\n\u003ctd style=\"width: 8.45254%; text-align: center;\"\u003e20ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 11.5689%; text-align: center;\"\u003eAc-DEVD-pNA (2 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 8.49085%; text-align: center;\"\u003e200ul\u003c\/td\u003e\n\u003ctd style=\"width: 8.45254%; text-align: center;\"\u003e1ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 11.5689%; text-align: center;\"\u003epNA (10 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 8.49085%; text-align: center;\"\u003e200ul\u003c\/td\u003e\n\u003ctd style=\"width: 8.45254%; text-align: center;\"\u003e1ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eWhen stored at-20 °C, Ac-DEVD-pNA and pNA need to be stored in the dark. Valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"20T","offer_id":41518131445835,"sku":"abs50025-20T","price":117.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41518131478603,"sku":"abs50025-100T","price":333.0,"currency_code":"USD","in_stock":true}]},{"product_id":"tunel-fitc","title":"TUNEL Assay Apoptosis Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e1. Experimental materials (self-prepared)\u003cbr\u003e\u003c\/strong\u003ePBS buffer (1 ×, pH ~ 7.4)\u003cbr\u003e0.4% Triton X-100 (PBS formulated)\u003cbr\u003e0.1% Triton X-100 in PBS containing 5 mg\/mL BSA\u003cbr\u003e4% Paraformaldehyde (PBS formulated)\u003cbr\u003eImmunohistochemical pen\u003cbr\u003eDewaxing Solvent (Paraffin Section Sample)\u003cbr\u003eReagents related to paraffin section processing\u003cbr\u003eAnti-fluorescence quenching sealed tablets\u003cbr\u003eddH\u003csub\u003e2\u003c\/sub\u003eO\u003cbr\u003e\u003cstrong\u003e2. Experimental design\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eA. Positive control:\u003cbr\u003e\u003c\/strong\u003ePositive control slides were prepared by DNase I treatment. DNase I can digest single-stranded or double-stranded DNA to expose the 3 '-OH terminus, artificially causing apoptosis. Just do it once per experiment. (Used to verify whether there are any problems with this experimental operation and kit)\u003cbr\u003e\u003cstrong\u003eB. Negative control:\u003cbr\u003e\u003c\/strong\u003eA TUNEL Reaction Buffer without TdT Enzyme was used to replace TdT Enzyme with ddH2O. (Mainly to exclude non-specific staining caused by cell apoptosis, operation process and other reasons; and to adjust the shooting exposure intensity.)\u003cbr\u003e\u003cstrong\u003eC. Experimental treatment group.\u003cbr\u003e\u003c\/strong\u003eThe experimental group operated normally according to the instructions.\u003cstrong\u003e\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eD. Experimental control group.\u003cbr\u003e\u003c\/strong\u003eThe experimental group operated normally according to the instructions.\u003cstrong\u003e\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e3. Experimental steps:\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e1. Sample preparation:\u003cbr\u003e\u003c\/strong\u003e(1) For adherent cells or cell smears\u003cbr\u003ea. PBS wash once.\u003cbr\u003eNote: If you are worried that the cells of the cell smear will not stick firmly, you can dry the sample to make the cells stick firmly.\u003cbr\u003eb. Fixation: Add an appropriate amount of 4% paraformaldehyde (formulated in PBS) and fix at 4 °C for 30 min. PBS wash twice.\u003cbr\u003ec. Permeability: Add an appropriate amount of 0.4% Triton X-100 (prepared in PBS), and permeate at room temperature for 20 minutes. PBS wash twice.\u003cbr\u003ed. Go to step 2. TUNEL reaction.\u003c\/p\u003e\n\u003cp\u003e(2) For suspended cells or cell suspensions\u003cbr\u003eA. Cells were collected (3-5 × 106 cells), centrifuged at 1000 rpm for 5 min, and washed twice with PBS.  \u003cbr\u003eb. Fixation: Add an appropriate amount of 4% paraformaldehyde (prepared with PBS) to fully resuspend the cells, and fix at 4 ℃ for 30 minutes. Centrifuge at 2000 rpm for 5 min and wash twice with PBS.  \u003cbr\u003ec. Permeability: Add an appropriate amount of 0.4% Triton X-100 (prepared in PBS), and permeate at room temperature for 20 minutes. Centrifuge at 2000 rpm for 5 min and wash twice with PBS.  \u003cbr\u003ed. Go to step 2. TUNEL reaction.  \u003c\/p\u003e\n\u003cp\u003e(3) Paraffin tissue section\u003cbr\u003eA. The paraffin sections were placed on a slicing rack and baked in an oven at 60 °C for 60 minutes.  \u003cbr\u003eb. Dewaxing and hydration: Section samples were sequentially placed into xylene I (10 min) → xylene II (10 min) → 100% ethanol I (5 min) → 100% ethanol II (5 min) → 95% ethanol (5 min) → 90% ethanol (5 min) → 80% ethanol (5 min) → 70% ethanol (5 min) → ddH2O rinse for 5 min and rinse twice.  \u003cbr\u003eNote: Xylene is toxic and volatile, please do this in a fume hood.  \u003cbr\u003ec. Use filter paper to dry the liquid around the section sample, and circle the sample outline with an immunohistochemical pen to facilitate downstream transparency and marking.  \u003cbr\u003eNote: If the outline circle of immunohistochemical strokes is found to be destroyed in subsequent experimental operations, it is necessary to make up the drawing in time.  \u003cbr\u003ed. Permeability: Dilute 2 mg\/mL Proteinase K solution with PBS at a ratio of 1: 50 to a final concentration of 40 µ g\/mL, add 100 µ L dropwise to each sample so that the solution covers the entire sample area and incubate at 37 °C for 30 min.  \u003cbr\u003eNote: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagent in the subsequent step can fully enter the nucleus for reaction, and improve the labeling efficiency. Too long incubation time will increase the risk of tissue sections falling off the carrier slice in subsequent washing steps, and too short incubation time may cause insufficient permeability treatment and affect labeling efficiency. In order to obtain better results, the concentration, incubation time and temperature of Proteinase K need to be optimized according to different types of tissue samples.  \u003cbr\u003ee. The slice samples were immersed in 1 × PBS and rinsed three times for 5 min each time, the excess liquid was aspirated with filter paper, and the processed samples were kept moist in a wet box.  \u003cbr\u003eNote: Proteinase K must be cleaned during this step, otherwise it will seriously interfere with the subsequent labeling reaction.  \u003cbr\u003ef. Go to step 2. TUNEL reaction.  \u003c\/p\u003e\n\u003cp\u003e(4) Frozen tissue section\u003cbr\u003eA. Fixation: Take out the frozen sections and warm them to room temperature. The slice samples were immersed in 4% paraformaldehyde (PBS formulated) and fixed at room temperature for 30 min. Section samples were immersed in 1 × PBS and rinsed three times for 10 min each.  \u003cbr\u003eNote: If you are worried that formaldehyde will not be cleaned clean, it will affect the final dyeing effect. After formaldehyde fixation is completed, an appropriate amount of 2 mg\/mL glycine can be added to wash for 10 minutes to neutralize the residual fixative, and then wash with PBS.  \u003cbr\u003eb. Use filter paper to dry the liquid around the section sample, and circle the sample outline with an immunohistochemical pen to facilitate downstream transparency and marking.  \u003cbr\u003eNote: If the outline circle of immunohistochemical strokes is found to be destroyed in subsequent experimental operations, it is necessary to make up the drawing in time.  \u003cbr\u003ec. Permeability: Dilute 2 mg\/mL Proteinase K solution with PBS to a final concentration of 40 µ g\/mL at a ratio of 1:50, add 100 µ L dropwise to each sample so that the solution covers the entire sample area and incubate at 37 ℃ for 20 min.  \u003cbr\u003eNote: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagent in the subsequent step can fully enter the nucleus for reaction, and improve the labeling efficiency. Too long incubation time will increase the risk of tissue sections falling off the carrier slice in subsequent washing steps, and too short incubation time may cause insufficient permeability treatment and affect labeling efficiency. In order to obtain better results, the concentration, incubation time and temperature of Proteinase K need to be optimized according to different types of tissue samples. If optimizing the concentration and incubation time of Proteinase K still cannot improve the staining effect, you can choose to immerse the sample in 1% Triton X-100 (prepared in PBS) at room temperature for 3-5 minutes; The slice samples were then immersed in 1 × PBS and rinsed three times for 5 min each.  \u003cbr\u003ed. The slice samples were immersed in 1 × PBS and rinsed three times for 5 min each time, the excess liquid was aspirated with filter paper, and the processed samples were kept moist in a wet box.  \u003c\/p\u003e\n\u003cp\u003e(5) Positive treatment (only positive control is subjected to this step, other samples are directly subjected to TUNEL reaction step)\u003cbr\u003eA. Dilute 10 × DNase I Buffer to 1 × DNase I Buffer with ddH20 at a ratio of 1: 10 for later use.\u003cbr\u003eb. Add 100 µ L of 1 × DNase I Buffer dropwise to the processed sample covering the entire sample area and equilibrate at room temperature for 5 min.\u003cbr\u003ec. DNase I (2 U\/μL) was diluted 1: 100 with 1 × DNase I Buffer to a final concentration of 20 U\/mL of the working solution.\u003cbr\u003ed. Discard the Buffer, add 100 μL of DNase I working solution at a concentration of 20 U\/mL, and incubate at room temperature for 15 min.\u003cbr\u003ee. Discard the DNase I working solution and wash twice with PBS.\u003cbr\u003ef. Go to step 2. TUNEL reaction.\u003cbr\u003e\u003cstrong\u003e2. TUNEL\u003c\/strong\u003e\u003cstrong\u003ereaction\u003cbr\u003e\u003c\/strong\u003e(1) Prepare TUNEL reaction solution (ready for use):\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 83.9685%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e1 sample\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e5 samples\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e10 samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003eTdT Enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003cp\u003e2 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\u003cp\u003e10 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\u003cp\u003e20 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e\u003cp\u003eTUNEL Reaction Buffer\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003cp\u003e48 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\u003cp\u003e240 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\u003cp\u003e480 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e\u003cp\u003eTotal TUNEL reaction solution volume\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003cp\u003e50 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\u003cp\u003e250 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\u003cp\u003e500 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e(2) For adherent cells, cell smears or tissue sections\u003cbr\u003eA. Add 50 μL of TUNEL reaction solution to each sample so that the reaction solution evenly covers the sample. Incubate at 37 ℃ in the dark for an appropriate time (the recommended staining time for cells is 15-30min, and the recommended staining time for tissues is 1 h).  \u003cbr\u003eNote: 50μL TUNEL reaction solution is suitable for smear, section or 96-well plate (other different well plates can appropriately adjust the volume of TUNEL reaction solution to cover cells). If the sample to be tested is a smear, section or in a 24-well plate, 12-well plate or 6-well plate, you can use an anti-evaporation film, or try to use a ziplock bag or other appropriate material to cut it into a round plastic sheet slightly smaller than the well. Add TUNEL reaction solution dropwise and cover the sample, which can prevent the evaporation of TUNEL reaction solution and make the TUNEL reaction solution evenly cover the sample.  \u003cbr\u003eb. Discard the TUNEL reaction solution, wash it twice with PBS, and then wash it three times with 0.1% Triton X-100 (formulated in PBS containing 5 mg\/mL BSA) for 5 minutes each time, so that free unreacted markers can be cleaned.  \u003cbr\u003e c. (Optional) Add an appropriate amount of DAPI dye solution with a concentration of 5μg\/mL to each sample, and incubate at room temperature and protect from light for 5 minutes. After completion of staining, discard the DAPI dye solution and wash it twice with PBS for 5 min each time.  \u003cbr\u003ed. (Optional) Section sealing: Add 20μL of anti-fluorescence quenching sealing tablet dropwise to each sample (anti-fluorescence quenching sealing tablet may not be suitable for some dyes, and it is recommended to conduct pre-experiment to test the matching before the experiment), cover the coverslip, and gently tap the coverslip with the blunt end of tweezers to remove air bubbles to complete the sealing.\u003cbr\u003ee. Aspirate the excess liquid with filter paper, add 100 μL of PBS to the sample area to keep the sample moist, and immediately observe under a fluorescence microscope.\u003cbr\u003e\u003cbr\u003e(3) For suspended cells or cell suspensions\u003cbr\u003eA. Add 50 μL of TUNEL reaction solution to each sample tube to gently resuspend cells, and incubate at 37 °C in the dark for 15-30 minutes. The cells were gently resuspended with a micropipette every 10 min.\u003cbr\u003eb. Centrifuge at 2000 rpm for 5 minutes, discard the TUNEL reaction solution, and wash twice with 0.1% Triton X-100 (PBS containing 5 mg\/mL BSA) for 5 minutes each time, so that free unreacted markers can be cleaned.  \u003cbr\u003ec. Each sample tube was added with 100 μL of DAPI dye solution with a concentration of 5 μg\/mL, and incubated at room temperature and protected from light for 5 minutes.\u003cbr\u003ed. 400 μL of PBS was added to resuspend cells and immediately detected by flow cytometry or smeared and observed under a fluorescence microscope.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTUNEL cell apoptosis detection kit; One-step TUNEL apoptosis detection kit (green fluorescence)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eSample types: paraffin sections, frozen sections, adherent cells and suspension cells.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eWhen cells undergo apoptosis, some DNA endonucleases will be activated. These endonucleases will cut off the genomic DNA between nucleosomes and produce DNA fragments of 180 bp to 200 bp, which shows the specific Ladder Ladder map presented in agarose gel electrophoresis. When genomic DNA double-stranded or single-stranded breaks, a large number of sticky 3 '-OH ends will be produced, which can be combined with 488\/Cy-dUTP under the catalysis of deoxyribonucleotide terminal transferase (TdT), so as to directly detect apoptotic cells through fluorescence microscope or flow cytometry. This method is called Terminal-deoxynucleotidyl transfer mediated nick end labeling (TUNEL). Since normal or proliferating cells have little DNA breakage, there is no 3 '-OH formation and little can be stained. Tunel method can stain complete single apoptotic nuclei or apoptotic bodies in situ, accurately reflect the typical biochemical and morphological characteristics of apoptosis, and can detect a very small number of apoptotic cells. Therefore, it is widely used in the research of apoptosis.  \u003cbr\u003eThe kit has a wide range of applications and can be used to detect the apoptosis of cells in frozen or paraffin sections, and can also detect the apoptosis of cultured adherent cells or suspended cells. It can selectively detect apoptotic cells, but not necrotic cells or cells with DNA strand breaks caused by irradiation and drug treatment. The kit takes short time to detect cell apoptosis, only needs one step of staining reaction, and can be detected after washing.\u003cbr\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 77.7072%; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eserial number\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e20T\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e50T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eTUNEL Reaction Buffer (488)\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e1 mL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e2 × 1. 25 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eTdT Enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e40 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e100 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eProteinase K (2 mg\/mL)\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e40 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e100 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eD\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eDNase I (2 U\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e5 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e13 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eE\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003e10 × DNase I Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e100 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e260 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Before use, please centrifuge the product to the bottom of the tube instantly, and then conduct subsequent experiments.  \u003cbr\u003e2. When the staining background is heavy or the non-specific staining is obvious, the staining time can be appropriately reduced.  \u003cbr\u003e3. It is recommended to add negative control and positive control group during the experiment.  \u003cbr\u003e4. Please wear a mask and gloves when using component A. If it comes into contact with skin, please rinse with plenty of water immediately.  \u003cbr\u003e5. Fluorescent dyes all have quenching problems. Please try to avoid light to slow down fluorescence quenching.  \u003cbr\u003e6. This product is limited to scientific research and cannot be used for clinical diagnosis or treatment, food or medicine, or stored in ordinary houses.  \u003cbr\u003e7. For your safety and health, please wear a lab coat and disposable gloves.  \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eWhen stored at-20 ℃, component A should be protected from light to avoid repeated freezing and thawing.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"20T","offer_id":41518136852555,"sku":"abs50047-20T","price":258.0,"currency_code":"USD","in_stock":true},{"title":"50T","offer_id":41518136885323,"sku":"abs50047-50T","price":442.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/74822de7afee4d7aae9c21d64f1cbd41.jpg?v=1789740048"},{"product_id":"edu-488-cell-proliferation-detection-kit","title":"EdU-488 Cell Proliferation Assay Kit (6-well plate)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eBefore the trial guidelines:\u003cbr\u003e1, the corresponding animal experiment, EdU suggested initial dosage of 50 mg\/kg (50 mu g\/g), dilute concentration is 0.5 1 mg\/mL. Are injected into mice weight 20 g, mg, 1 to 1 mg\/mL, need injection of 1 mL.\u003cbr\u003e2, for the cells, the recommended 10 microns, EdU work for in vitro culture cell, specific EdU use concentration, incubation time along with the samples and research purpose is different, can be adjusted appropriately.\u003cbr\u003e3. This kit can be used for EdU experiments of about 200T in 6-well plates, and EdU experiments of about 2000T in 96-well plates or paraffin sections. The amount of use should be calculated according to the sample conditions during the experiment.\u003cbr\u003eI. Sample processing\u003cbr\u003e1. Animal experiments\u003cbr\u003e(1) EdU injection in animals\u003cbr\u003eFor mice, EdU can be prepared with PBS at a dose of 10 to 200mg\/kg and injected intraperitoneally, locally into specific tissues or organs, or added to drinking water. The specific dosage is related to the type, weight and mode of use of animals, and can be referred to the relevant literature. Therefore, it is recommended to conduct a certain exploration of the use concentration of EdU for the first use, or directly use the concentration of 50mg\/kg for testing.\u003cbr\u003eInjection methods: according to customer experiments, such as intraperitoneal injection, subcutaneous injection, intramuscular injection, tail vein injection, etc., among which intraperitoneal injection is the most common.\u003cbr\u003eAfter 6 hours or according to the specific experiments to determine the appropriate time, quickly kill mice, take out the organization, in accordance with the general steps to make frozen section and paraffin section. EdU tag when also can consult relevant literature to adjust.\u003cbr\u003eSmall intestinal epithelial cells proliferate rapidly, and positive information can be detected 6 hours after EdU injection in adult mice, which can be used as a positive control for pre-experiments.\u003cbr\u003e(2) slice processing\u003cbr\u003eBefore slicing processing: best cleaning tissues and organs, to remove residues in the tissues of the blood, EdU, reduce the background.\u003cbr\u003eIt is advisable to slice thickness: 3-10 microns, biopsy had thick could affect the background and dyeing efficiency.\u003cbr\u003eSlice post-processing:\u003cbr\u003eParaffin dewaxing: dimethyl benzene dewaxing 2 times, 15 minutes per times, gradient ethanol (100%, 95%, 85%, 75%), 5 minutes per times, deionized water elution 1 times.\u003cbr\u003eProcessing of frozen sections: After 30 min at room temperature, the sections were fixed for 10 min and washed three times with PBS for 5 min each.\u003cbr\u003e2, cell experiment\u003cbr\u003e(1) EdU labeling of cells\u003cbr\u003ea.with cell complete medium and EdU mother liquor by 2000:1 the ratio of dilution, preparation of a moderate amount of 100 microns EdU medium;\u003cbr\u003eb.each hole adding suitable amount of 10 microns EdU medium 2 hours incubation, abandon culture medium (optimal incubation time for 1\/10 of the cell cycle) commonly, EdU incubation medium and EdU reaction liquid volume may refer to the table below;\u003cbr\u003ec.Cells were washed 3 times with PBS for 5 minutes each time\u003cbr\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 97.1447%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 17.054%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 11.598%; text-align: center;\"\u003e96-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e48-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e24-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e12-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e6-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e5.5cm capsule\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 17.054%; text-align: center;\"\u003eEdU medium\u003c\/td\u003e\n\u003ctd style=\"width: 11.598%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 17.054%; text-align: center;\"\u003eEdU reaction solution\u003c\/td\u003e\n\u003ctd style=\"width: 11.598%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e(2) cells fixed fully\u003cbr\u003ea.Cell fixative (PBS containing 4% paraformaldehyde) was added to each well and incubated at room temperature for 15 minutes. The fixative was discarded and washed with PBS for 3 times, 5 minutes each time.\u003cbr\u003eb.Each well was decolorized with an osmotic agent (0.5% TritonX-100 in PBS) and incubated for 15 minutes. PBS was washed three times for 5 min each.\u003cbr\u003e\u003cbr\u003eSecond, EdU response\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 40.4358%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 46.1517%; height: 22px; text-align: center;\"\u003eConstituent\u003c\/td\u003e\n\u003ctd style=\"width: 50.4245%; height: 22px; text-align: center;\"\u003eDosage\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 46.1517%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 50.4245%; height: 22px; text-align: center;\"\u003e855μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 46.1517%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 50.4245%; height: 22px; text-align: center;\"\u003e40μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 46.1517%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 50.4245%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 46.1517%; height: 22px; text-align: center;\"\u003e488 primary color\u003c\/td\u003e\n\u003ctd style=\"width: 50.4245%; height: 22px; text-align: center;\"\u003e5μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 46.1517%; height: 22px; text-align: center;\"\u003eReaction liquid volume\u003c\/td\u003e\n\u003ctd style=\"width: 50.4245%; height: 22px; text-align: center;\"\u003e1000μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003eThe EdU reaction solution was prepared according to the ratio of PB: CU: AC: 488 chromogen = 855:40:100:5 (the reaction solution was prepared on the spot).   \u003cbr\u003eadd 50-500\u0026amp;mu per sample dropfold; L EdU dyeing liquid evenly cover samples (reaction), reaction liquid at room temperature away from light incubation 15 to 60 minutes, abandon the dyeing liquid reaction, PBS wash three times, each time for 5 minutes.   \u0026lt; br \/ \u0026gt; 3, DAPI staining \u0026amp; have spent \u0026lt; br \/ \u0026gt; DAPI (100 x), with PBS according to the scale of 1:100 dilution into ready-to-use DAPI staining fluid, each sample in 50-500 \u0026amp; mu; L DAPI staining solution, staining in the dark for 15 min, and washing with PBS three times for 5 min each. \u0026lt; br \/ \u0026gt; 4, image photos \u0026amp; have spent \u0026lt; br \/ \u0026gt; dyeing advice immediately after the completion of observation, using fluorescence microscopy, confocal microscope or film scanner to capture images, after dyed dark glass slides 4 ℃. \u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231113\/cc27d047061c4da09106133ba29cc2e5.png\" alt=\"\" width=\"806\" height=\"466\"\u003e\u003cbr\u003eAbove are stained images of 50mg\/kg labeled mice for 6 hours\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eEdU(5-ethynyl-2' -deoxyuridine), also known as 5-ethynyl-2' -deoxyuridine (5-ethynyl-2' -deoxyuridine), is a thymidine analogue with an alkynyl hydroxyl group that is rare in natural compounds and can replace thymidine (thymidine, 5-ethynyl-2' -deoxyuridine) in cell proliferation. thymidine (thymidine) is inserted into the replicating DNA molecule, the ethynyl group on EdU can react covalently with the fluorescently labeled small molecule Azide probe (Azide Alexa Fluor 488, etc.) through the catalysis of copper monovalent ion to form a stable triazole ring. \"It is called the Clickreaction, which allows for an efficient and rapid measurement of cell proliferation, especially the percentage of cells in S phase.\" Compared with the traditional immunofluorescence staining (BrdU) detection method, EdU is only 1\/500 of the size of BrdU antibody, and it is easier to diffuse in cells. It does not require strict sample denaturation (acid hydrolysis, pyolysis, and enzymatic hydrolysis), which effectively avoids sample damage and helps to observe the real situation of cell proliferation at the overall level of tissues and organs. Has higher sensitivity and faster detection speed.\u003cbr\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 65.4028%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003eNumber\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003eSize\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003eStorage Temp.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e120mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e1.7g\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e4.4mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e488 primary color\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e550μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e5\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e100mg\/mL EdU mother solution\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003eAccording to PB, CU: AC: 488 chromogen = 855: spoken, Catalan than column configuration EdU reaction liquid (liquid of reaction is used).\u003cbr\u003eEach sample add 50-500 mu L EdU staining reaction liquid liquid evenly cover samples (reaction), 15 to 60 minutes at room temperature away from light incubation, abandon the dyeing liquid reaction, PBS wash 3 times, each time for 5 minutes.\u003cbr\u003e3. DAPI staining\u003cbr\u003eDAPI(100x) was diluted with PBS at a ratio of 1:100 to make ready-to-use DAPI staining solution. 50-500μL of DAPI staining solution was added to each sample and stained in the dark for 15 minutes, followed by three washes with PBS for 5 minutes each.\u003cbr\u003e4. Take pictures\u003cbr\u003eFluorescence microscope, confocal microscope or whole-film scanner were used to collect images. After staining, the slides were stored in a dark place at 4℃.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231113\/cc27d047061c4da09106133ba29cc2e5.png\" alt=\"\" width=\"806\" height=\"466\"\u003e\u003cbr\u003eAbove are stained images of 50mg\/kg labeled mice for 6 hours.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e-20 ° C, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"200T","offer_id":41518137442379,"sku":"abs50050-200T","price":440.0,"currency_code":"USD","in_stock":true}]},{"product_id":"edu-555-cell-proliferation-detection-kit","title":"EdU-555 cell proliferation assay kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eBefore the trial guidelines\u003cbr\u003e1, the corresponding animal experiment, EdU suggested initial dosage of 50 mg\/kg (50 mu g\/g), dilute concentration is 0.5 1 mg\/mL. Are injected into mice weight 20 g, mg, 1 to 1 mg\/mL, need injection of 1 mL.\u003cbr\u003e2, for the cells, the recommended 10 microns, EdU work for in vitro culture cell, specific EdU use concentration, incubation time along with the samples and research purpose is different, can be adjusted appropriately.\u003cbr\u003e3, this kit can be carried out about 200 t of 6 orifice EdU experiments, about 2000 t96 orifice or paraffin section EdU experiment, experiment should be calculated according to the sample situation in the process of usage.\u003cbr\u003eI. Sample processing\u003cbr\u003e1, the animal experiment\u003cbr\u003e(1) EdU injection in animals\u003cbr\u003eIn mice, can according to the amount of 10-200 mg\/kg, the EdU with PBS mixed with a certain concentration, intraperitoneal injection, specific tissues or organs local injections or added to drinking water. The specific dosage is related to the type, weight and mode of use of animals, and can be referred to the relevant literature. Therefore, it is recommended to conduct a certain exploration of the use concentration of EdU for the first use, or directly use the concentration of 50mg\/kg for testing.\u003cbr\u003eInjection method: the experiment according to the customer, such as abdominal cavity injection, subcutaneous injection, intramuscular, tail intravenous injection, etc., among them with intraperitoneal injection.\u003cbr\u003eAfter 6 h or after the appropriate time was determined according to the specific experiment, the mice were quickly sacrificed, the desired tissue was removed, and frozen or paraffin sections were made according to routine procedures. EdU labeling can also be adjusted by referring to the relevant literature.\u003cbr\u003eSmall intestinal epithelial cells proliferate rapidly, and positive information can be detected 6 hours after EdU injection in adult mice, which can be used as a positive control for pre-experiments.\u003cbr\u003e(2) Slice processing\u003cbr\u003ePre-section treatment: It is best for tissues and organs to be washed to remove residual EdU in blood and tissues and reduce background.\u003cbr\u003eSection thickness: 3-10μm is appropriate, too thick section may affect the section background and staining efficiency.\u003cbr\u003eSlice post-processing:\u003cbr\u003eParaffin sections were deparaffinized twice with xylene for 15 min, once each with gradient ethanol (100%, 95%, 85%, 75%) for 5 min, and once with deionized water for elution.\u003cbr\u003eFrozen section processing: room temperature placed after 30 minutes, fixed 10 minutes, PBS cleaning three times, each time for 5 minutes.\u003cbr\u003e2. Cell experiments\u003cbr\u003e(1) cells EdU markers\u003cbr\u003ea.with cell complete medium and EdU mother liquor by 2000:1 the ratio of dilution, preparation of a moderate amount of 10 microns EdU medium;\u003cbr\u003eb.each hole adding suitable amount of 10 microns EdU medium 2 hours incubation, abandon culture medium (optimal incubation time for 1\/10 of the cell cycle) commonly, EdU incubation medium and EdU reaction liquid volume may refer to the table below;\u003cbr\u003ec.PBS cleaning cell 3 times, each time for 5 minutes.\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 94.6731%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e96-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e48-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e24-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e12-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e6-well plates\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e5.5cm capsule\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003eEdU medium\u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 16.0584%; text-align: center;\"\u003eEdU reaction solution\u003c\/td\u003e\n\u003ctd style=\"width: 12.5936%; text-align: center;\"\u003e50μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.6534%; text-align: center;\"\u003e70μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.5444%; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 12.3756%; text-align: center;\"\u003e200μL\u003c\/td\u003e\n\u003ctd style=\"width: 10.9433%; text-align: center;\"\u003e500μL\u003c\/td\u003e\n\u003ctd style=\"width: 11.0712%; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e(2) cells fixed fully\u003cbr\u003ea. Cell fixative (PBS containing 4% paraformaldehyde) was added to each well and incubated at room temperature for 15 minutes. The fixative was discarded and washed with PBS for 3 times, 5 minutes each time.\u003cbr\u003eb. Each well was decolorized with an osmotic agent (0.5% TritonX-100 in PBS) and incubated for 15 minutes. PBS was washed three times for 5 min each.\u003cbr\u003e\u003cbr\u003eSecond, EdU response\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 50%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eConstituent\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eDosage\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e855μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e40μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e555 primary colour\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e5μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003eReaction liquid volume\u003c\/td\u003e\n\u003ctd style=\"width: 48.2881%; height: 22px; text-align: center;\"\u003e1000μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003eThe EdU reaction solution was prepared according to the ratio of PB: CU: AC: 555 chromogen = 855:40:100:5 (the reaction solution was prepared immediately).\u003cbr\u003eEach sample add 50-500 mu L EdU staining reaction liquid liquid evenly cover samples (reaction), 15 to 60 minutes at room temperature away from light incubation, abandon the dyeing liquid reaction, PBS wash 3 times, each time for 5 minutes.\u003cbr\u003e3. DAPI staining\u003cbr\u003eDAPI(100x) was diluted with PBS at a ratio of 1:100 to make ready-to-use DAPI staining solution. 50-500μL of DAPI staining solution was added to each sample and stained in the dark for 15 minutes, followed by three washes with PBS for 5 minutes each.\u003cbr\u003e4. Take pictures\u003cbr\u003eFluorescence microscope, confocal microscope or whole-film scanner were used to collect images. After staining, the slides were stored in a dark place at 4℃.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20231114\/5d82c6d2133649808e68f720411fd044.png\" alt=\"\" width=\"752\" height=\"528\"\u003e\u003cbr\u003eAbove are stained images of 50mg\/kg labeled mice for 6 hours\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eEdU(5-ethynyl-2' -deoxyuridine), also known as 5-ethynyl-2' -deoxyuridine (5-ethynyl-2' -deoxyuridine), is a thymidine analogue with an alkynyl hydroxyl group that is rare in natural compounds and can replace thymidine (thymidine, 5-ethynyl-2' -deoxyuridine) in cell proliferation. thymidine (thymidine) is inserted into the replicating DNA molecule, the ethynyl group on EdU can react covalently with the fluorescently labeled small molecule Azide probe (Azide Alexa Fluor 488, etc.) through the catalysis of copper monovalent ion to form a stable triazole ring. \"It is called the Clickreaction, which allows for an efficient and rapid measurement of cell proliferation, especially the percentage of cells in S phase.\" With traditional immunofluorescence staining (BrdU) method compared with EdU only 1\/500 the size of the BrdU antibody, are more likely to spread within the cell, don't need strict sample degeneration (acid solution, pyrolysis, enzyme solution) processing, effectively avoid the damage samples, help in the overall level of tissue and organ observation of cell proliferation, It has higher sensitivity and faster detection speed.Note: AC is a powder, need to add 10mL pure water before use, fully dissolved before use, AC is easy to oxidation failure, after the refrigerator is stored at -20 degrees, if found discoloration, need to replace fresh reagent. In animal experiments, the mass concentration of 100mM EdU mother liquor corresponds to 25mg\/mL, which should be converted according to the dosage.\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 65.4028%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003eNO.\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003eSize\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003eStorage Temp.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003ePB\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e120mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eAC\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e1.7g\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e3\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003eCU\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e4.4mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e555 Primary Colour\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e550μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 12.6323%; height: 22px; text-align: center;\"\u003e5\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; height: 22px; text-align: center;\"\u003e100mg\/mL EdU mother solution\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; height: 22px; text-align: center;\"\u003e100μL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 12.6323%; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003ctd style=\"width: 38.4719%; text-align: center;\"\u003eDAPI（100×）\u003c\/td\u003e\n\u003ctd style=\"width: 17.0952%; text-align: center;\"\u003e1.2mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.7639%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003eNote: AC is a powder, need to add 10mL of pure water before use, fully dissolved before use, AC is easy to oxidation failure, after packaging and stored in the refrigerator -20 degrees, if found discoloration, need to replace fresh reagent. In animal experiments, the mass concentration of 100mM EdU mother liquor corresponds to 25mg\/mL, which should be converted according to the dosage.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e-20 ° C, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"200T","offer_id":41518137999435,"sku":"abs50051-200T","price":440.0,"currency_code":"USD","in_stock":true}]},{"product_id":"calcein-am-pi-double-stain-kit","title":"Calcein-AM\/PI Double Stain Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e1. Preparation of working fluid:\u003c\/strong\u003e\u003cbr\u003e1. Preparation of 1 × Assay Buffer\u003cbr\u003eTake out 10 × Assay Buffer from low-temperature refrigerator, take out an appropriate amount according to the aseptic conditions of single dosage, and dilute 10 times with deionized water (dH2O) to obtain 1 × Assay Buffer.\u003cbr\u003e2. Preparation of 1 × dyeing working solution\u003cbr\u003e(1) First, return the cryopreserved Calcein-AM solution (2mM) and PI solution (1.5 mM) to room temperature for 20-30min.\u003cbr\u003eNote: The mother liquor can be dispensed for the first use to reduce the number of repeated freezing and thawing.\u003cbr\u003e(2) Add 5μl of Calcein-AM solution (2mM) and 15μl of PI solution (1.5 mM) to 5ml of 1 × Assay Buffer and mix well. At this time, the working solution concentration of Calcein-AM was 2 μM, and the working solution concentration of PI was 4.5 μM. Because the optimal staining conditions of different cell lines are different, it is recommended to do gradient experiments for the initial experiment to determine the optimal concentrations of Calcein-AM and PI. The principle of gradient screening is to use the lowest probe concentration to obtain the best fluorescence results.\u003cbr\u003e\u003cstrong\u003eNote: Due to the poor stability of Calcein-AM, this dyeing solution must be prepared immediately and used up the same day.\u003c\/strong\u003e\u003cbr\u003e\u003cstrong\u003e2. Dyeing steps:\u003c\/strong\u003e\u003cbr\u003e1. For adherent cells, first digest the cells with a cell spatula or trypsin-EDTA, and then collect the cells by centrifugation (1000rpm, 3min). For suspended cells, cells were collected by direct centrifugation (1000 rpm, 3 min).\u003cbr\u003e2. Remove the supernatant and fully wash the cells 2 to 3 times with 1 × Assay Buffer to fully remove the residual esterase activity.\u003cbr\u003e3. Prepare the cell suspension with 1 × Assay Buffer to have a density of 1 × 105 ~ 1 × 106 cells\/ml.\u003cbr\u003e4. Take 100μl of staining working solution and add it to 200μl of cell suspension, mix well, and incubate at 37 °C for 15min.\u003cbr\u003e\u003cstrong\u003eNote: Incubation time can be extended to 30min if desired.\u003c\/strong\u003e\u003cbr\u003e5. Use a 490 ± 10 nm excitation filter under a fluorescence microscope to simultaneously detect living cells (yellow-green fluorescence) and dead cells (red fluorescence). In addition, only dead cells were observed using a 545 nm emission filter. Detection can also be performed directly under a fluorescent microplate reader using a suitable filter.\u003cbr\u003e\u003cstrong\u003eNote: The following methods can be used to optimize the optimum working concentrations for the two fluorescent dyes.\u003cbr\u003e\u003c\/strong\u003e(1) incubating the cells with 0.1% saponin or 0.1-0.5% digoxin for 10 minutes, or incubating the cells with 70% ethanol for 30 minutes, thereby preparing dead cells;\u003cbr\u003e(2) Dead cells were stained with 0.1-10 μM PI solution to obtain the optimal working concentration for staining only the nucleus, but not the cytoplasm.\u003cbr\u003e(3) Dead cell staining was performed with 0.1-10 μM Calcein-AM to obtain an optimal working concentration that would not stain the cytoplasm. Live cells are then stained at this concentration to see if the live cells can be stained\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCalcein-AM\/PI\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCalcein-AM is a cell staining reagent that fluorescently labels living cells and fluoresces green (Ex = 490nm, Em = 515nm). Because it introduces acetylmethoxymethyl ester (AM) group on the basis of traditional Calcein, it increases hydrophobicity and makes it easily penetrate living cell membranes. Once in the cell, Calcein-AM (which itself does not fluoresce) is cleaved by esterases in the cell to form a membrane-impermeable polar molecule Calcein, which is retained in the cell and emits strong green fluorescence. Compared with other similar reagents (such as BCECF-AM and CFDA), Calcein, AM is the most suitable fluorescent probe for staining viable cells due to its extremely low cytotoxicity, and does not inhibit any cellular functions, such as proliferation and lymphocyte chemotaxis.\u003cbr\u003eBecause dead cells lack esterase, Calcein-AM is only used for cell viability testing and short-term labeling of living cells. Therefore, Calcein-AM is often used in combination with dead cell fluorescent probes such as propidium iodide (PI) to perform fluorescent double staining of living cells and dead cells at the same time. Propidium iodide (PI) cannot pass through the cell membrane of living cells, but can only pass through the disordered area of the dead cell membrane to reach the nucleus, and embedded into the DNA double helix of the cell to produce red fluorescence (Ex = 535 nm, Em = 617 nm), so PI only stains dead cells. Since both Calcein and PI-DNA can be excited at 490 nm, live and dead cells can be observed simultaneously with fluorescence microscope. With 545 nm excitation, only dead cells can be observed.\u003cbr\u003eThe working principle of this kit lies in the double dyes of Calcein-AM and PI to perform double staining labeling of living cells and dead cells, so as to analyze the level of living cells and dead cells. According to our optimized experimental system, a single staining of 200μl cell suspension can perform 500 tests.\u003cbr\u003eProduct Components:\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 89.384%; height: 87px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.4578%; height: 22px; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 32.9926%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 13.7317%; height: 22px; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd style=\"width: 32.7187%; height: 22px; text-align: center;\"\u003eSave\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 13.4578%; height: 21px; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 32.9926%; height: 21px; text-align: center;\"\u003eCalcein-AM Solution (2mM)\u003c\/td\u003e\n\u003ctd style=\"width: 13.7317%; height: 21px; text-align: center;\"\u003e50 μL\u003c\/td\u003e\n\u003ctd style=\"width: 32.7187%; height: 21px; text-align: center;\"\u003eStore dry at-20 ℃ protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.4578%; height: 22px; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 32.9926%; height: 22px; text-align: center;\"\u003ePI Solution (1.5 mM)\u003c\/td\u003e\n\u003ctd style=\"width: 13.7317%; height: 22px; text-align: center;\"\u003e150 μL\u003c\/td\u003e\n\u003ctd style=\"width: 32.7187%; height: 22px; text-align: center;\"\u003eStore dry at-20 ℃ protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 13.4578%; height: 22px; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 32.9926%; height: 22px; text-align: center;\"\u003e10 × Assay Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 13.7317%; height: 22px; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 32.7187%; height: 22px; text-align: center;\"\u003eStore at-20 °C, and can be stored at 4 °C for regular use.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Since Calcein-AM is very sensitive to humidity, if the required amount of Calcein-AM solution is taken every time, the lid must be tightly sealed. It is recommended to package and seal storage according to the single dosage. The Calcein-AM working fluid must be prepared for ready use.\u003cbr\u003e2. Propidium iodide (PI) has certain carcinogenicity, so we must pay attention to protection during operation. If it comes into contact with the skin, it needs to be washed with tap water immediately.\u003cbr\u003e3. For your safety and health, please wear a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Preparation of working solution: \u003cbr\u003e1, 1× The preparation of Assay Buffer (reaction buffer) \u003cbr\u003eRemove from the low temperature refrigerator for 10 times; Assay Buffer, according to the single dosage of sterile conditions to take out the appropriate amount, with deionized water (dH2O) to do 10 times dilution to get 1times; Assay Buffer. \u003cbr\u003e2, 1× Dyeing the configuration of the working liquid \u003cbr\u003e(1) the Calcein - AM solution to be kept at low temperature (2 mM) and PI solution (1.5 mM) back to the room temperature 20-30 min. \u003cbr\u003eNote: The mother liquor can be divided for the first use to reduce the number of repeated freezing-thawing. \u003cbr\u003e(2) apply 5 \u0026amp; mu; l Calcein-AM solution (2mM) and 15μ l PI solution (1.5mM) added to 5ml 1× Assay Buffer and mix well. At this time, the working solution concentration of Calcein-AM obtained is 2μ M, PI working liquid concentration is 4.5 \u0026amp; mu; M. As the optimal staining conditions differ for different cell lines, gradient experiments are recommended for initial experiments to determine the optimal concentration of Calcein-AM and PI. The principle of gradient screening is to use the lowest probe concentration to obtain the best fluorescence results. \u003cbr\u003e\u003cstrong\u003eNote: Due to the poor stability of Calcein-AM, this staining solution must be used on the go and used up the same day. \u003c\/strong\u003e\u003cbr\u003e\u003cstrong\u003eII. Staining steps: \u003c\/strong\u003e\u003cbr\u003e1. For adherent cells, first digest the cells with a cell scraper or pancrease-EDTA, and then collect the cells by centrifugation (1000rpm, 3min). For suspended cells, cells were collected by centrifugation directly (1000 rpm, 3min). \u003cbr\u003e2, remove the supernatant and use 1× The cells were thoroughly washed 2 to 3 times with Assay Buffer to fully remove residual esterase activity. \u003cbr\u003e3, with 1× The cell suspension was prepared with Assay Buffer to a density of 1× 105~1× 106 cells \/ml. \u003cbr\u003e4, take 100μ l Add 200\u0026amp;mu to staining working solution; l cell suspension was mixed and incubated at 37 ° C for 15min. \u003cbr\u003e\u003cstrong\u003eNote: The incubation time can be extended to 30min if desired. \u003c\/strong\u003e\u003cbr\u003e5, use 490\u0026amp;plusmn under fluorescence microscope; 10 nm excitation filter was used to detect both live cells (yellow-green fluorescence) and dead cells (red fluorescence). In addition, only dead cells could be observed using the 545nm emission filter. It can also be detected directly under a fluorescent microplate reader using a suitable filter. \u003cbr\u003e\u003cstrong\u003eNote: The following methods can be used to optimize the optimal working concentration of the two fluorescent dyes. \u003cbr\u003e\u003c\/strong\u003e(1) Dead cells were prepared by incubating cells with 0.1% saponin or 0.1-0.5% digoxin for 10min or 70% ethanol for 30min; \u003cbr\u003e(2) use 0.1-10μ M's PI solution was used for dead cell staining to obtain an optimal working concentration that stained only the nucleus, but not the cytoplasm. \u003cbr\u003e(3) with 0.1-10μ M of Calcein-AM was used for dead cell staining to obtain an optimal working concentration that would not stain the cytoplasm. This concentration was then used for live-cell staining to see if viable cells could be stained\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eStored dry at-20 ℃ in the dark from light, the shelf life is 12 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"500T","offer_id":41518138556491,"sku":"abs50056-500T","price":137.0,"currency_code":"USD","in_stock":true}]},{"product_id":"live-cell-tracking-kit","title":"Living cells tracer kit (green fluorescence)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSelf-brought consumables:\u003cbr\u003e\u003c\/strong\u003e24 well plate, adjustable pipette gun and tip\u003cbr\u003eCentrifuge\u003cbr\u003eFluorescence microscope or flow cytometry\u003cbr\u003eDeionized water, PBS\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eReagent Preparation:\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eCell Tracker Green\u003c\/strong\u003e\u003cstrong\u003e：\u003c\/strong\u003eOperate on ice, protect from light and store in separate packages to avoid repeated freezing and thawing.\u003cbr\u003e\u003cstrong\u003e1 × Assay Buffer\u003c\/strong\u003e\u003cstrong\u003e：\u003c\/strong\u003eDilute 5 × Assay Buffer to 1 × Assay Buffer with deionized water. Heat to 37 °C before use.\u003cbr\u003e\u003cstrong\u003eStaining Solution\u003c\/strong\u003e\u003cstrong\u003e：\u003c\/strong\u003ePrior to the start of the experiment, the Cell Tracker Green was diluted 1: 1000 to 1 × Assay Buffer and the dose was expanded accordingly for large sample size determination. Protect from light and preheat to 37 °C before use.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eattention\u003c\/strong\u003e\u003cstrong\u003e:\u003c\/strong\u003e\u003cstrong\u003eFor different cell types and\u003c\/strong\u003e\u003cstrong\u003e\/\u003c\/strong\u003e\u003cstrong\u003eOr experimental conditions, which should be determined empirically\u003c\/strong\u003e\u003cstrong\u003eCell Tracker Green\u003c\/strong\u003e\u003cstrong\u003eThe final concentration of. Typically, long-term staining (more than about\u003c\/strong\u003e\u003cstrong\u003e3\u003c\/strong\u003e\u003cstrong\u003eDays) or using rapidly dividing cells would require a 1: 1000 dilution to double the dye concentration. For shorter experiments (e.g. viability assays), it may be necessary to use concentrations as low as 1:\u003c\/strong\u003e\u003cstrong\u003e5000\u003c\/strong\u003e\u003cstrong\u003eDiluted dye. In order to maintain a normal cellular physiological state and reduce potential artifacts, the concentration of the dye should be kept as low as possible.\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure:\u003cbr\u003e\u003c\/strong\u003eA. Flow cytometry quantification\u003cbr\u003e1. Treat the cells with the required method and establish a parallel control group;\u003cbr\u003eNote: Parallel control suggestions: unstained cells (i.e. cells without Cell Tracker Green), suspended with 1 × Assay Buffer;\u003cbr\u003e2. For non-adherent cells, 0.5-1x10 were collected by centrifugation (4 °C, 300 g, 5 min)\u003csup\u003e6\u003c\/sup\u003eCells. Wash twice with pre-cooled PBS and discard the PBS. For adherent cells, cells were first digested with trypsin (without EDTA) and then centrifuged;\u003cbr\u003e3. Resuspending the cell pellet in 500 uL Staining Solution;\u003cbr\u003e4. Incubate cells at 37 °C for 15-30 min under dark conditions:\u003cbr\u003e5. Isolate the cells at 500 g and discard the supernatant;\u003cbr\u003e6. Resuspend the pellet in fresh preheated medium, incubate the cells for another 30 min to ensure complete contact with the probe, and wash the cells with PBS again;\u003cbr\u003e7. At 5x10\u003csup\u003e5\u003c\/sup\u003eTo 1x10\u003csup\u003e6\u003c\/sup\u003eDensity of cells\/ml Cell pellets were weighted in preheated PBS and cells were immediately analyzed by flow cytometry using an FL1 channel (typically FL1).\u003c\/p\u003e\n\u003cp\u003eB. Fluorescence microscope detection\u003cbr\u003e1. Make a fine sound on the stem: Perform flow cytometry according to steps A.1 to A.7, and place the cell burst of A.7 on slide 1. The cells were beautified with a glass coverslip. Cells were analyzed as soon as possible by fluorescence microscopy using an appropriate filter. If the cells are to be fixed and permeabilized, proceed to step B.3.\u003cbr\u003e2. For adherent cells: the recommended protocol is as follows\u003cbr\u003e(1) Cells were directly cultured with 24-well culture blood. CO at 37 °C before staining\u003csub\u003e2\u003c\/sub\u003eCultivate in an incubator for at least 24 h;\u003cbr\u003e(2) treating the cells with a desired method;\u003cbr\u003e(3) washing the cells twice with PBS and discarding the PBS;\u003cbr\u003e(4) 0.5 mL of Staining Solution was added to white blood cells and incubated at 37 ° C. for 15-30 min in the dark;\u003cbr\u003e(5) Discard the supernatant, replace it with preheated fresh growth medium, and then incubate at 37 ° C. for another 30 min in the dark;\u003cbr\u003e(6) Cells were washed twice with PBS or an appropriate buffer. If the cells are to be fixed and permeabilized, proceed to step B.3;\u003cbr\u003e(7) Use a suitable filter for observation.\u003cbr\u003e3. Fixation and permeabilization\u003cbr\u003e(1) Fixation with an aldehyde-containing fixative. Usually we used 4% paraformaldehyde (abs9179) to fix cells at room temperature for 15 min;\u003cbr\u003e(2) After fixation, the cells were washed with PBS three times;\u003cbr\u003e(3) After fixation, if the cells are to be detected with antibodies later, they should be incubated in ice-cold acetone for 10 minutes to make the cells permeable. After permeation, the cells should be washed 3 times with PBS.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eThe study of cell movement and localization requires the use of specific probes that are non-toxic to living cells. Live Cell Tracking Kit provides a versatile and well-preserved cell tracking reagent (Cell Tracker Green) for monitoring cell movement, location, proliferation, migration, chemotaxis, and invasiveness. Cell Tracker Green is a live cell fluorescent tracer probe that can enter cells through passive diffusion and covalently bind to intracellular proteins. It is a long-acting cell tracer dye. Once in the cell, the non-fluorescent Cell Tracker Green will be hydrolyzed by endolesterase to produce green fluorescence (Ex\/Em = 494\/521 nm). These fluorescent products can only accumulate in cells with intact cell membranes, so dead cells and intact cell membranes cannot be stained. Cell Tracker Green is not sensitive to pH changes and can be fixed by formaldehyde or glutaraldehyde. The Cell Tracker Green probe can be retained in living cells for several generations and can show fluorescence for at least one week. The probe can be transferred to daughter cells, but not to adjacent cells in the population.\u003cbr\u003eProduct Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 70.7692%; height: 66px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 43.535%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 21.8727%; height: 22px; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd style=\"width: 29.3305%; height: 22px; text-align: center;\"\u003eStorage conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 43.535%; height: 22px; text-align: center;\"\u003eCell Tracker Green\u003c\/td\u003e\n\u003ctd style=\"width: 21.8727%; height: 22px; text-align: center;\"\u003e250uL\u003c\/td\u003e\n\u003ctd style=\"width: 29.3305%; height: 22px; text-align: center;\"\u003e-20 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 43.535%; height: 22px; text-align: center;\"\u003eAssay Buffer (5 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 21.8727%; height: 22px; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 29.3305%; height: 22px; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003eProduct Features:\u003cbr\u003eUsed to monitor cell motility, localization, proliferation, migration, chemotaxis and invasion.\u003cbr\u003eThe CellTracker Green probe can be well retained in living cells for several generations and can show fluorescence for at least one week. The probe is transferred to daughter cells, but not to neighboring cells in the population.\u003cbr\u003eOptimized for flow cytometry or fluorescence microscopy.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBackground\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCell motility and localization studies require specialized probes that must be non-toxic to living cells. Living cell tracer kit (green fluorescence) provides convenience for monitoring cell movement, localization, proliferation, migration, chemotaxis and invasion.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eStore at-20 ℃ in the dark from light, shelf life is 12 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"500T","offer_id":41518139736139,"sku":"abs50060-500T","price":250.0,"currency_code":"USD","in_stock":true}]},{"product_id":"cell-migration-assay-kit","title":"Cell Migration Assay Kit (24-well plate, 8μM)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSelf-brought instruments, reagents and consumables:\u003cbr\u003e\u003c\/strong\u003eMigratory cell line\u003cbr\u003eCell culture medium with 10% fetal calf serum\u003cbr\u003eSerum-free medium (i.e. cell culture medium with 0% fetal bovine serum)\u003cbr\u003eCell incubator (37 ℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e）\u003cbr\u003ePBS, cotton swab, forceps, 96-well plate, 24-well plate, adjustable pipette gun and tip\u003cbr\u003eMicroscope\u003cbr\u003eMicroplate reader (capable of measuring absorbance at 570 nm)\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eReagent Preparation:\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e24-Well Migration Plate\u003c\/strong\u003e\u003cstrong\u003e：\u003c\/strong\u003eReady-to-use; Before use, equilibrate to room temperature; Store at 4 °C.\u003cbr\u003e\u003cstrong\u003eFixation\u003c\/strong\u003e\u003cstrong\u003e：\u003c\/strong\u003eReady-to-use; Before use, equilibrate to room temperature; Store at-20 °C.\u003cbr\u003e\u003cstrong\u003eStaining Solution\u003c\/strong\u003e\u003cstrong\u003e：\u003c\/strong\u003eReady-to-use; Before use, equilibrate to room temperature; Store at 4 °C.\u003cbr\u003e\u003cstrong\u003eElution Solution\u003c\/strong\u003e\u003cstrong\u003e：\u003c\/strong\u003eReady-to-use; Before use, equilibrate to room temperature; Store at 4 °C.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure:\u003c\/strong\u003e\u003cstrong\u003eNote: Before the formal experiment, be sure to conduct a pre-experiment to determine the best experimental conditions for cell migration, such as cell number, cell culture time and staining time.\u003cbr\u003e\u003c\/strong\u003e1. In an ultra-clean workbench, the 24-well Migration Plate is equilibrated at room temperature for 10 minutes.\u003cbr\u003e2. Dilute the cells to 0.5-5.0 × 10 with serum-free medium\u003csup\u003e6\u003c\/sup\u003eCells\/mL to prepare a cell suspension.\u003cbr\u003e\u003cstrong\u003eNote: It is recommended that cells be starved overnight before cell migration.\u003cbr\u003e\u003c\/strong\u003e3. Add 600 µ L of culture medium containing 10% fetal bovine serum or the required chemoattractant to the lower chamber, then add the small chamber to wet, and add 100 µ L of cell suspension to the upper chamber.\u003cbr\u003e\u003cstrong\u003eNote: (\u003c\/strong\u003e\u003cstrong\u003e1\u003c\/strong\u003e\u003cstrong\u003e) The solution in the upper and lower chambers must not produce bubbles, so the migration will be weakened; (\u003c\/strong\u003e\u003cstrong\u003e2\u003c\/strong\u003e\u003cstrong\u003e) Cell status is critical, and try to choose cells with fewer passages.\u003cbr\u003e\u003c\/strong\u003e4. In the cell incubator (37 ℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e) for 2-24 h.\u003cbr\u003e\u003cstrong\u003eNote: Too long culture time may cause some migrated cells to fall off from the bottom surface of the membrane. It is recommended to select the appropriate culture time through pre-experiment.\u003cbr\u003e\u003c\/strong\u003e5. Carefully take out the small chamber, suck out the culture medium in the upper chamber, and gently wipe off the unmigrated cells in the upper chamber with the end of a wet cotton swab.\u003cbr\u003e\u003cstrong\u003eNote: Wipe off the unmigrated cells in the upper chamber. The action must be gentle and do not puncture the polycarbonate membrane.\u003cbr\u003e\u003c\/strong\u003e6. Transfer that chamber to a well containing 600 uL of Fixation Solution, fix at room temperature for 10 min, and wash 3 times with PBS.\u003cbr\u003e7. Then transfer the chamber to 600L Staining Solution, stain at room temperature for 5-15 minutes, wash 3 times with PBS, 3 minutes each time, and dry. (If the chamber is still dark stained, it is recommended to gently wash the chamber 3 times in a beaker filled with PBS and dry).\u003cbr\u003e\u003cstrong\u003eNote: Both ambient temperature and reagent temperature will affect the dyeing time, but a shorter dyeing time will only affect the depth of coloring. If the color is lighter, the dyeing time can be properly heated or extended to achieve the required dyeing effect. Dip dyeing at 37 ℃ for 15 minutes can ensure full coloring.\u003cbr\u003e\u003c\/strong\u003e8. Place the chamber in a clean hole, select at least 3 visual fields with the microscope to observe and take photos, and calculate the number of migrating cells.\u003cbr\u003e9. After taking photos, add 400 µ L of Elution Solution to the lower chamber, elute for 10 minutes, completely elute the Staining Solution, aspirate 200 µ L of the eluted solution into a 96-well plate, and measure the OD value at 570 nm.\u003cbr\u003e\u003cstrong\u003eNote: (1) Usually, it takes 10 minutes to complete elution at a room temperature of 22-28C. The elution time can be appropriately adjusted according to the temperature difference. (2) You can choose staining photos and measure OD values according to specific experimental needs.\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCell migration, generally speaking, refers to the characteristic of cells migrating from one place to another when stimulated by foreign signals, usually during processes such as wound healing, cell differentiation, embryonic development and tumor metastasis. Transwell is an experimental technology that can simulate many mucosal and biological barrier systems of the body in vitro. The main material of this technology is Transwell chamber. The cell migration analysis kit (24-well plate, 8 µ m) uses polycarbonate membrane (8 µ m pore size) to determine the migration characteristics of cells. The principle is to put the Transwell chamber into a culture plate, the chamber is called the upper chamber, and the culture plate is called the lower chamber. The upper and lower culture medium are separated by polycarbonate membrane.\u003cbr\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 76.54%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 45.5955%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 22.1365%; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd style=\"width: 26.9326%; text-align: center;\"\u003eStorage conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 45.5955%; text-align: center;\"\u003e24-Well Migration Plate\u003c\/td\u003e\n\u003ctd style=\"width: 22.1365%; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 26.9326%; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 45.5955%; text-align: center;\"\u003eFixation\u003c\/td\u003e\n\u003ctd style=\"width: 22.1365%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.9326%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 45.5955%; text-align: center;\"\u003eStaining Solution\u003c\/td\u003e\n\u003ctd style=\"width: 22.1365%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.9326%; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 45.5955%; text-align: center;\"\u003eElution Solution\u003c\/td\u003e\n\u003ctd style=\"width: 22.1365%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 26.9326%; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003eProduct Features:\u003cbr\u003eEasy and quick to operate.\u003cbr\u003eSimple, optimized experimental method.\u003cbr\u003eThe migration ability of the cells was tested.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eOwn instruments, reagents, consumables: \u003cbr\u003e\u003c\/strong\u003eMigratory cell lines \u003cbr\u003eCell culture medium with 10% fetal bovine serum \u003cbr\u003eserum-free medium (i.e. cell culture medium with 0% fetal bovine serum) \u003cbr\u003eCell culture incubator (37℃, 10 ℃), 5% CO\u003csub\u003e2\u003c\/sub\u003e\u003cbr\u003ePBS \u003cbr\u003e, cotton swabs, forceps, 96-well plate, 24-well plate, adjustable pipette gun and tip \u003cbr\u003emicroscope \u003cbr\u003emicroplate reader (can measure the absorbance at 570 nm) \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eReagent preparation: \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e24-Well Migration Plate\u003c\/strong\u003e\u003cstrong\u003e: \u003c\/strong\u003eread-to-use; Before use, balance to room temperature; Store at 4 ° C. \u003cbr\u003e\u003cstrong\u003eFixation Solution\u003c\/strong\u003e\u003cstrong\u003e: \u003c\/strong\u003eread-to-use type; Before use, balance to room temperature; Store at -20 ° C. \u003cbr\u003e\u003cstrong\u003eStaining Solution\u003c\/strong\u003e\u003cstrong\u003e: \u003c\/strong\u003eread-to-use; Before use, balance to room temperature; Store at 4 ° C. \u003cbr\u003e\u003cstrong\u003eElution Solution\u003c\/strong\u003e\u003cstrong\u003e: \u003c\/strong\u003eready-to-use; Before use, equilibrate to room temperature; Store at 4 ° C. \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure: \u003c\/strong\u003e\u003cstrong\u003eNote: Before the formal experiment, it is important to conduct a pre-experiment to determine the best experimental conditions for cell migration, such as cell number, cell culture time, and staining time. \u003cbr\u003e\u003c\/strong\u003e1, in an ultra-clean workbench, the 24-well Migration Plate was equilibrated at room temperature for 10 min. \u003cbr\u003e2, cells were diluted to 0.5-5.0\u0026amp;times with serum-free medium; 10\u003csup\u003e6\u003c\/sup\u003ecells \/mL, cell suspension was prepared. \u003cbr\u003e\u003cstrong\u003eNote: It is recommended to starve the cells overnight before doing cell migration. \u003cbr\u003e\u003c\/strong\u003e3. Add 600 \u0026amp;micro to the lower chamber; L medium containing 10% fetal bovine serum or the desired chemoattractant, then add chamber wetting, add 100 \u0026amp;micro to the upper chamber; L of cell suspension. \u003cbr\u003e\u003cstrong\u003eNote: (\u003c\/strong\u003e\u003cstrong\u003e1\u003c\/strong\u003e\u003cstrong\u003e) The solution in the upper and lower chambers must not produce bubbles, so that the migration effect will be weakened; (\u003c\/strong\u003e\u003cstrong\u003e2\u003c\/strong\u003e\u003cstrong\u003e) Cell status is critical, try to choose cells with fewer passages. \u003cbr\u003e\u003c\/strong\u003e4. The cells were cultured in a cell incubator (37℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e) for 2-24 hours. \u003cbr\u003e\u003cstrong\u003eNote: Too long culture time may cause some migrating cells to fall off from the bottom surface of the membrane, so it is recommended to choose the appropriate culture time by pre-experiment. \u003cbr\u003e\u003c\/strong\u003e5, carefully remove the chamber, suck out the medium in the upper chamber, and gently wipe off the non-migrated cells in the upper chamber with the end of a wet cotton swab. \u003cbr\u003e\u003cstrong\u003eNote: Wipe off the upper chamber non-migrated cells, the action must be gentle, do not Pierce the polycarbonate membrane. \u003cbr\u003e\u003c\/strong\u003e6, the chamber was transferred to a well containing 600 uL Fixation Solution, fixed at room temperature for 10 min, and washed 3 times with PBS. \u003cbr\u003e7, the chamber was then transferred to the well containing 600L Staining Solution, stained at room temperature for 5-15 min, washed 3 times with PBS for 3 min each time, and allowed to dry. (If the chamber still has dark staining, it is recommended to gently clean the chamber in a beaker with PBS for 3 times and let it dry). \u003cbr\u003e\u003cstrong\u003eNote: Both ambient temperature and reagent temperature will affect the time required for staining, but a short staining time will only affect the depth of the color. If the color is light, heating or prolonging the staining time can be used to achieve the desired staining effect, and dyeing at 37℃ for 15 min can ensure sufficient staining. \u003cbr\u003e\u003c\/strong\u003e8. The chamber was placed in a clean well, and at least 3 fields of view were selected for observation and photo taking under the microscope, and the number of migrating cells was calculated. \u003cbr\u003e9, after taking photos, add 400 \u0026amp;micro to the lower chamber; L Elution Solution, elution for 10 min, the Staining Solution was completely eluted, and the eluted solution was absorbed 200   µ L into a 96-well plate and OD was measured at 570 nm. \u003cbr\u003e\u003cstrong\u003eNote: (1) Usually in 22-28C room temperature environment, it takes 10 min to complete the elution, the elution time can be adjusted according to the temperature difference. (2) The staining can be selected to take photos and measure OD value according to the specific experimental requirements. \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eStore it separately according to the prompt of each component label, and the validity period is 6 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"12T","offer_id":41518139965515,"sku":"abs50061-12T","price":533.0,"currency_code":"USD","in_stock":true}]},{"product_id":"cell-invasion-assay-kit","title":"Cell Invasion Assay kit (24-well plate, 8μM)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSelf-brought instruments, reagents and consumables:\u003cbr\u003e\u003c\/strong\u003eInvasive cell line\u003cbr\u003eCell culture medium with 10% fetal calf serum\u003cbr\u003eSerum-free medium (i.e. cell culture medium with 0% fetal bovine serum)\u003cbr\u003eCell incubator (37 ℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e）\u003cbr\u003ePBS, cotton swab, forceps, 96-well plate, 24-well plate, adjustable pipette gun and tip\u003cbr\u003eMicroscope\u003cbr\u003eMicroplate reader (capable of measuring absorbance at 570 nm)\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eReagent Preparation:\u003cbr\u003e\u003c\/strong\u003e24-Well Invasion Plate: ready-to-use; Before use, equilibrate to room temperature; Store at 4 °C.\u003cbr\u003eMatrigel: thaw at 4 °C; Place it on ice before use, and store the unused reagents at-80 ℃ in divided packs.\u003cbr\u003eFixation Solution: ready to use; Before use, equilibrate to room temperature; Store at-20 °C.\u003cbr\u003eStaining Solution: ready to use; Before use, equilibrate to room temperature; Store at 4 °C.\u003cbr\u003eElution Solution: Ready-to-use; Before use, equilibrate to room temperature; Store at 4 °C.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure:\u003cbr\u003eNote: Before the formal experiment, a pre-experiment must be conducted to determine the best experimental conditions for cell invasion. Cells must have the ability to secrete matrix metalloproteinases (MMPs) in order to undergo cell invasion. It is recommended to conduct a qPCR pre-experiment to detect the expression of MMPs, especially MMP-2, before the experiment.\u003cbr\u003e\u003c\/strong\u003e1. Prepare Matrigel: Put the gun tip, gun tip box, EP tube, serum-free medium and 24-Well Invasion Plate into the refrigerator for pre-cooling. The Matrigel was removed from − 80 °C, and the Matrigel was thawed at 4 °C. After thawing, the Matrigel was placed in an ice box for operation experiment.\u003cbr\u003e\u003cstrong\u003eNote: Before the Matrigel forms a gel, mix well with a pre-cooled sterile tip and the Matrigel melts into a clear liquid.\u003cbr\u003e\u003c\/strong\u003e2. Dilute Matrigel: Dilute Matrigel with pre-cooled serum-free medium in an ultra-clean workbench.\u003cbr\u003e\u003cstrong\u003eNote: (1) Select the appropriate dilution ratio of Matrigel according to the experimental conditions. The dilution ratio setting range is Matrigel: serum-free medium = 1: 6-1: 8, and the recommended dilution ratio is 1: 6. (2) It is not recommended to keep unused Matrigel after dilution for reuse.\u003cbr\u003e\u003c\/strong\u003e3. Please draw 45 µ L of diluted Matrigel from the pre-cooled sterile tip, add it to the upper chamber of 24-Well Invasion Plate, spread it evenly at the bottom, and put it in the cell incubator (37 ℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e) for 1 h to polymerize Matrigel into gel. According to the actual effect of gel film formation, the incubation time can be appropriately extended.\u003cbr\u003e\u003cstrong\u003eNote: Do not produce bubbles during the glue laying process, and lay the glue evenly. Generally, the gel can be incubated for 1 hour. It is difficult to see clearly after the gel, so the next experiment can be done directly.\u003cbr\u003e\u003c\/strong\u003e4. Suck off the excess liquid in the upper chamber, add 100 µ L of serum-free medium to each well, and place it in the incubator for 30 minutes to hydrate the basement membrane.\u003cbr\u003e5. Suck off the liquid in the upper chamber, check that the liquid has not passed through the small chamber and entered the lower chamber, and then seed the cells.\u003cbr\u003e\u003cstrong\u003eNote: If liquid enters the lower chamber, the experimental results may be inaccurate. It is recommended to re-lay the glue.\u003cbr\u003e\u003c\/strong\u003e6. Dilute the cells to 0.5-5.0 × 10 with serum-free medium\u003csup\u003e6\u003c\/sup\u003eCells\/mL to prepare a cell suspension.\u003cbr\u003e\u003cstrong\u003eNote: It is recommended that cells be starved overnight before cell invasion.\u003cbr\u003e\u003c\/strong\u003e7. Add 600 µ L of culture medium containing 10% fetal bovine serum or the required chemoattractant to the lower chamber, then add the small chamber to wet, and add 100 µ L of cell suspension to the upper chamber.\u003cbr\u003e\u003cstrong\u003eNote: (1) The solution in the upper and lower chambers must not produce bubbles, so that the invasion will be weakened; (2) Cell status is very critical. Try to choose cells with fewer passages.\u003cbr\u003e\u003c\/strong\u003e8. In the cell incubator (37 ℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e) for 12-48 h.\u003cbr\u003e\u003cstrong\u003eNote: Too long culture time may cause some invaded cells to fall off from the bottom surface of the membrane. It is recommended to select the appropriate culture time through pre-experiment.\u003cbr\u003e\u003c\/strong\u003e9. Carefully take out the small chamber, suck out the culture medium in the upper chamber, and gently wipe off the uninvaded cells in the upper chamber with the end of a wet cotton swab.\u003cbr\u003e\u003cstrong\u003eNote: Wipe off the uninvaded cells in the upper chamber. The movement must be gentle and do not puncture the polycarbonate membrane.\u003cbr\u003e\u003c\/strong\u003e10. Transfer the chamber to a well containing 600 µ L of Fixation Solution, fix at room temperature for 10 min, and wash 3 times with PBS.\u003cbr\u003e11. Transfer the chamber to a well containing 600 µ L Staining Solution, stain at room temperature for 5-15 min, wash 3 times with PBS for 3 min each time, and air dry. (If the chamber is still dark stained, it is recommended to gently wash the chamber 3 times in a beaker filled with PBS and dry).\u003cbr\u003e\u003cstrong\u003eNote: Both ambient temperature and reagent temperature will affect the dyeing time, but a shorter dyeing time will only affect the depth of coloring. If the color is lighter, the dyeing time can be properly heated or extended to achieve the required dyeing effect. Dip dyeing at 37 ℃ for 15 minutes can ensure full coloring.\u003cbr\u003e\u003c\/strong\u003e12. Place the chamber in a clean hole, select at least 3 visual fields with the microscope to observe and take pictures, and calculate the number of invading cells.\u003cbr\u003e13. After taking photos, add 400 µ L of Elution Solution to the lower chamber, elute for 10 minutes, completely elute the Staining Solution, aspirate 200 µ L of the eluted solution into a 96-well plate, and measure the OD value at 570 nm.\u003cbr\u003e\u003cstrong\u003eNote: (1) Usually, it takes 10 minutes to complete the elution at a room temperature of 22-28 °C. The elution time can be appropriately adjusted according to the temperature difference. (2) You can choose staining photos and measure OD values according to specific experimental needs.\u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eCell invasiveness refers to the characteristic that cells are stimulated by foreign signals and invade from one place to another. It usually occurs in processes such as wound healing, cell differentiation, embryonic development and tumor metastasis. Transwell is an experimental technology that can simulate many mucosal and biological barrier systems in vitro. The main material of this technology is Transwell chamber. The cell invasion analysis kit (24-well plate, 8 µ m) uses polycarbonate membrane (8 µ m pore size) to determine the invasion characteristics of cells. Matrigel (Matrigel) is the basement membrane matrix extracted from EHS mouse tumors rich in extracellular matrix proteins. Its main components are laminin, type IV collagen, nestin, heparin glycoprotein, and also contains growth factors and matrix metalloproteinases. The principle is to put the Transwell chamber into the culture plate, the chamber is called the upper chamber, and the culture plate is called the lower chamber. The upper and lower layers of culture medium are separated by a polycarbonate membrane, and the matrigel is laid on the upper chamber of the membrane to simulate the extracellular matrix in vivo. If the cells in the upper chamber are to be transferred to the lower chamber, matrix metalloproteinases (MMPs) need to be secreted to degrade the matrigel. Because of the permeability of the polycarbonate membrane, the components in the lower culture medium can affect the cells in the upper chamber. By staining and elution, the number of cells is counted to reflect the invasive ability of cells.\u003cbr\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"http:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin_new_bak-english\/20260126\/b2f57b57ed01403c84f5a28471127440.jpg\" alt=\"\" width=\"472\" height=\"171\"\u003e\u003cbr\u003e\u003cstrong\u003eProduct Components:\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 64.2201%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49.9959%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 16.9144%; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd style=\"width: 27.7543%; text-align: center;\"\u003eStorage conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49.9959%; text-align: center;\"\u003e24-Well Invasion Plate\u003c\/td\u003e\n\u003ctd style=\"width: 16.9144%; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 27.7543%; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49.9959%; text-align: center;\"\u003eFixation\u003c\/td\u003e\n\u003ctd style=\"width: 16.9144%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 27.7543%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49.9959%; text-align: center;\"\u003eStaining Solution\u003c\/td\u003e\n\u003ctd style=\"width: 16.9144%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 27.7543%; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49.9959%; text-align: center;\"\u003eElution Solution\u003c\/td\u003e\n\u003ctd style=\"width: 16.9144%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 27.7543%; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 49.9959%; text-align: center;\"\u003eMatrigel\u003c\/td\u003e\n\u003ctd style=\"width: 16.9144%; text-align: center;\"\u003e100 µ L\u003c\/td\u003e\n\u003ctd style=\"width: 27.7543%; text-align: center;\"\u003e-80℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cstrong\u003e\u003cbr\u003eProduct Features:\u003c\/strong\u003e\u003cbr\u003eEasy and quick to operate.\u003cbr\u003eSimple, optimized experimental method.\u003cbr\u003eThe invasive ability of the cells was tested.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eOwn instruments, reagents, consumables: \u003cbr\u003e\u003c\/strong\u003eInvasive cell lines \u003cbr\u003eCell culture medium with 10% fetal bovine serum \u003cbr\u003eserum-free medium (i.e. cell culture medium with 0% fetal bovine serum) \u003cbr\u003eCell culture incubator (37 ° C, 5% CO\u003csub\u003e2\u003c\/sub\u003e\u003cbr\u003ePBS \u003cbr\u003e, cotton swabs, forceps, 96 well plate, 24 well plate, adjustable pipet gun and tip \u003cbr\u003emicroscope \u003cbr\u003emicroplate reader (can measure the absorbance at 570 nm) \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003ereagent preparation: \u003cbr\u003e\u003c\/strong\u003e24-Well Invasion Plate: ready-use type; Before use, balance to room temperature; Store at 4 ° C. \u003cbr\u003eMatrigel: Thaw at 4 ° C; Place on ice before use, and unusable reagents are stored in aliquots at -80 ° C. \u003cbr\u003eFixation Solution: ready-to-use type; Before use, balance to room temperature; Store at -20 ° C. \u003cbr\u003eStaining Solution: ready-to-use; Before use, balance to room temperature; Store at 4 ° C. \u003cbr\u003eElution Solution: ready-to-use type; Before use, equilibrate to room temperature; Store at 4 ° C. \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure: \u003cbr\u003eNote: Before the formal experiment, it is important to perform a pre-experiment to determine the optimal experimental conditions for cell invasion. Cells must have the ability to secrete matrix metalloproteinases (MMPs) in order to carry out cell invasion. It is recommended to perform qPCR pre-experiments to detect the expression of MMPs, especially MMP-2, before experiments. \u003cbr\u003e\u003c\/strong\u003e1. Prepare the matrix glue: Place the gun tip, gun tip box, EP tube, serum-free medium and 24-Well Invasion Plate in the refrigerator for pre-cooling. The Matrigel was removed from -80℃ and thawed at 4℃. After thawing, the Matrigel was placed in an ice box to operate the experiment. \u003cbr\u003e\u003cstrong\u003eNote: Before the formation of Matrigel gel, mix the whole process with a precooled sterile tip. After the Matrigel melts, it is a clear liquid. \u003cbr\u003e\u003c\/strong\u003e2. Dilute Matrigel: Dilute Matrigel with pre-cooled serum-free medium in a super-clean bench. \u003cbr\u003e\u003cstrong\u003eNote: (1) Select the appropriate dilution ratio of Matrigel according to the experimental conditions. The dilution ratio is set to Matrigel: serum-free medium =1:6-1:8, and the recommended dilution ratio is 1:6. (2) The unused Matrigel after dilution is not recommended to be retained for reuse. \u003cbr\u003e\u003c\/strong\u003e3, precooled sterile gun tip suction 45 µ The diluted Matrigel was added to the upper chamber of 24-Well Invasion Plate, evenly spread on the bottom, and placed in the cell culture incubator (37℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e) for 1 h to polymerize Matrigel into a gel. According to the actual effect of gel film formation, The incubation time can be extended appropriately. \u003cbr\u003e\u003cstrong\u003eNote: Do not produce bubbles during the spreading process, spreading the glue evenly. Generally, the gel can be incubated for 1 h. It is difficult to see after the gel, so the next experiment can be done directly. \u003cbr\u003e\u003c\/strong\u003e4. Absorb the excess liquid in the upper chamber and add 100\u0026amp;micro to each well; After L serum-free medium, the cells were placed in the incubator for 30 min, and the basement membrane was hydrated. \u003cbr\u003e5, the liquid in the upper chamber is sucked out, and after checking that the liquid does not pass through the chamber into the lower chamber, the cells are inoculated. \u003cbr\u003e\u003cstrong\u003eNote: If there is liquid into the lower chamber, the experimental results may not be accurate, it is recommended to re-spread the glue. \u003cbr\u003e\u003c\/strong\u003e6. Dilute the cells to 0.5-5.0\u0026amp;times with serum-free medium; 10\u003csup\u003e6\u003c\/sup\u003ecells \/mL, cell suspension was prepared. \u003cbr\u003e\u003cstrong\u003eNote: Cells are recommended to be starved overnight before cell invasion. \u003cbr\u003e\u003c\/strong\u003e7. Add 600 \u0026amp;micro to the lower chamber; L medium containing 10% fetal bovine serum or the desired chemoattractant, then add chamber wetting, add 100 \u0026amp;micro to the upper chamber; L of cell suspension. \u003cbr\u003e\u003cstrong\u003eNote: (1) The solution of the upper and lower chambers must not produce bubbles, so that the invasion effect will be weakened; (2) The state of cells is critical, try to choose cells with fewer passages. \u003cbr\u003e\u003c\/strong\u003e8. The cells were cultured in a cell incubator (37℃, 5% CO\u003csub\u003e2\u003c\/sub\u003e) for 12-48 hours. \u003cbr\u003e\u003cstrong\u003eNote: Too long culture time may cause some invasive cells to fall off from the bottom surface of the membrane, so it is recommended to choose the appropriate culture time by pre-experiment. \u003cbr\u003e\u003c\/strong\u003e9, carefully remove the chamber, suck out the medium in the upper chamber, and gently wipe off the non-invaded cells in the upper chamber with the end of a wet cotton swab. \u003cbr\u003e\u003cstrong\u003eNote: To wipe off the non-invaded cells in the upper chamber, the action must be gentle and do not Pierce the polycarbonate membrane. \u003cbr\u003e\u003c\/strong\u003e10, transfer the chamber to contain 600 µ L Fixation Solution hole, fixed 10 min at room temperature \u0026amp; have spent , PBS \u0026amp; have spent Wash 3 times. \u003cbr\u003e11, to transfer the small room to contain 600 \u0026amp; micro; L Staining Solution Wells were stained at room temperature for 5-15 min, washed 3 times with PBS for 3 min each, and allowed to dry. (if the small room still has deep color, suggested in a beaker of PBS gently cleaning chamber 3 \u0026amp; have spent Allow to dry). \u003cbr\u003e\u003cstrong\u003enote: the environment temperature and reagent temperature will affect the time needed for dyeing, but short dyeing time affect the depth of the color only, if the color shallow but appropriate heating or extend the dyeing time to achieve the required effect, see at 37 ℃ for 15 min ensures full color. \u003cbr\u003e\u003c\/strong\u003e12, the chamber was placed in a clean well, and at least 3 fields of view were selected for observation and photography under the microscope, and the number of invasive cells was calculated. \u003cbr\u003e13, after taking photos, add 400 \u0026amp;micro to the lower chamber; L Elution Solution and Elution 10 min, the Staining Solution Elution down completely, absorbs the Elution Solution after 200 \u0026amp; micro; L into a 96-well plate and OD was measured at 570 nm. \u003cbr\u003e\u003cstrong\u003enote: (1) usually in 22 to 28 ℃ under room temperature environment, need 10 min to complete elution, can be adjusted according to the temperature difference appropriate elution time. (2) The staining can be selected to take photos and measure OD value according to the specific experimental requirements. \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eStore it separately according to the prompt of each component label, and the validity period is 6 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"12T","offer_id":41518140293195,"sku":"abs50062-12T","price":600.0,"currency_code":"USD","in_stock":true}]},{"product_id":"senescence-b-galactosidase-staining-kit","title":"Cell senescence Assay Kit (β-galactosidase assay)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSelf-brought instruments, reagents and consumables:\u003c\/strong\u003e\u003cbr\u003eMicroscope\u003cbr\u003ewithout CO\u003csub\u003e2\u003c\/sub\u003e37 °C incubator\u003cbr\u003ePipette and tip, polypropylene tube (15 or 50 mL)\u003cbr\u003eVarious glassware for preparing reagents and buffer solutions\u003cbr\u003eDeionized water, glycerol\u003cbr\u003eSealing film, 6-well plate\u003cbr\u003e\u003cstrong\u003eReagent Preparation:\u003cbr\u003e\u003c\/strong\u003e1 × PBS: Before use, dilute to 1 × PBS with deionized water and equilibrate to room temperature; Store at 4 °C.\u003cbr\u003e1 × Fixation Buffer: Before use, dilute to 1 × Fixation Buffer with 1 × PBS and equilibrate to room temperature; Store at-20 °C.\u003cbr\u003eReagent A: ready-to-use; Before use, equilibrate to room temperature; Store at-20 °C.\u003cbr\u003eReagent B: ready-to-use; Before use, equilibrate to room temperature; Store at-20 °C.\u003cbr\u003eReagent C: ready-to-use; Before use, equilibrate to room temperature; Store at-20 °C.\u003cbr\u003eX-Gal solution: ready-to-use; Before use, equilibrate to 37 °C; Store at-20 °C protected from light. It is very important to heat the X-gal solution at 37 °C for 1 h to avoid the formation of aggregates, which may interfere with the visualization of stained cells.\u003cbr\u003eStaining Working Solution: Press 10 µ L Reagent A, 10 µ L Reagent B, 10 µ L Reagent C, 50 µ L X-gal solution And 920 µ L of 1 × PBS, then adjust to pH 6.0 with NaOH or HCl.\u003cbr\u003e\u003cstrong\u003eNote: When preparing the dyeing solution, please use containers made of polypropylene material or glass containers. A small amount of flocculation precipitation may occur and it will dissolve completely after shaking and mixing. Make sure it dissolves completely before use.\u003cbr\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure:\u003cbr\u003eNote: Staining Working Solution is toxic and corrosive to the human body. Please be careful when handling and pay attention to effective protection. Avoid contact with human body or direct inhalation.\u003c\/strong\u003e\u003cbr\u003e1. For adherent cells:\u003cbr\u003e1. Cultivate the cells directly on the coverslip of a 6-well plate and perform the required treatment.\u003cbr\u003e2. Aspirate the cell culture medium and wash it twice with 1 × PBS.\u003cbr\u003e3. Add 1 mL of 1 × Fixation Buffer to each well and fix at room temperature for 15 minutes. For other types of culture plates, the amounts of fixative and subsequent solutions are operated according to this ratio.\u003cbr\u003e4. Aspirate the cell fixative and wash the cells 3 times with 1 × PBS.\u003cbr\u003e5. Aspirate 1 × PBS and add 1 mL of Staining Working Solution to each well.\u003cbr\u003e6. Without CO\u003csub\u003e2\u003c\/sub\u003eIncubate overnight in a 37 °C incubator until the cells stained blue.\u003cbr\u003e\u003cstrong\u003eNote: Seal the 6-well plate with a sealing film to prevent cells from drying out and select the appropriate staining time. Cells cannot be cultured in an incubator containing CO2 because cell senescence staining is pH-related\u003c\/strong\u003e。\u003cbr\u003e7. Observe the cells under an optical microscope. The number of blue cells and total cells was counted, and the percentage of cells expressing beta-galactosidase (senescent cells) was calculated. After staining, if the counting cannot be observed in time, the Staining Working Solution can be removed, 2 mL of 1 × PBS can be added, and the cells can be stored at 4 ℃ for several days or covered with 70% glycerol solution for long-term storage at 4 ℃.\u003cbr\u003e\u003cbr\u003e2. For suspension cells:\u003cbr\u003e1. Centrifuge at 1,000 rpm for 5 min. Collect cells into 1.5 mL centrifuge tubes and wash twice with 1 × PBS.  \u003cbr\u003e2. Add 1 mL of 1 × Fixation Buffer to each tube and fix at room temperature for 15 minutes. When fixed, it can be shaken slowly on the shaker to avoid the cells forming clumps.\u003cbr\u003e3. At 1,000 rpm, centrifuge for 5 minutes, aspirate the cell fixative, and wash the cells 3 times with 1 × PBS.\u003cbr\u003e4. Aspirate 1 × PBS and add 1 mL of Staining Working Solution to each tube.\u003cbr\u003e5. Without CO\u003csub\u003e2\u003c\/sub\u003eIncubate overnight in a 37 °C incubator until the cells stained blue.\u003cbr\u003e6. Take part of the stained cells, add them dropwise to a glass slide or a 6-well plate, and observe under an ordinary optical microscope. The number of blue cells and total cells was counted, and the percentage of cells expressing beta-galactosidase (senescent cells) was calculated. If the counting cannot be observed in time, the Staining Working Solution can be removed, 2 mL of 1 × PBS can be added, and the cells can be stored at 4 °C for several days or covered with 70% glycerol solution for long-term storage at 4 °C.\u003c\/p\u003e\n\u003cp\u003e3. For tissue sections:\u003cbr\u003eThe paraffin sections were first dewaxed and hydrated according to conventional methods. Follow the following steps directly for frozen sections.\u003cbr\u003e1. For the prepared tissue sections, add an appropriate volume of 1 × Fixation Buffer to fully cover the tissue, and fix it at room temperature for 15mim\u003cbr\u003e2. Soak and wash the tissue with 1 × PBS for 3 times.\u003cbr\u003e3. Aspirate 1 × PBS and add an appropriate amount of Staining Working Solution.\u003cbr\u003e4. Without CO\u003csub\u003e2\u003c\/sub\u003eIncubate overnight in a 37 °C incubator until the cells stained blue.\u003cbr\u003e5. Observe the tissue sections under an optical microscope. The number of blue cells and total cells was counted, and the percentage of cells expressing beta-galactosidase (senescent cells) was calculated. After staining, if it cannot be observed in time, cover the tissue sections with 70% glycerol solution and store them at 4 °C for a long time.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eCell is the basic unit of the structure and function of organisms, and it is also the basic unit of aging of organisms. Cell senescence is morphologically manifested as degenerative changes in cell structure, such as nuclear membrane depression, which eventually leads to nuclear membrane disintegration, chromatin structure changes, and an increase in the number of hyperdiploid and abnormal polyploid cells; The fragility of cell membrane increases, the selective permeability decreases, and the type, number and sensitivity of membrane receptors to ligands change; Lipofuscin accumulates in cells, and many organelles and intracellular structures undergo degenerative changes. The physiological manifestations of cell senescence are functional decline and low metabolism, such as cell cycle arrest, loss of cell replication ability, weakened responsiveness to mitogenic stimuli, and changes in responsiveness to pro-apoptotic factors; Intracellular enzyme active centers are oxidized, enzyme activity decreases, protein synthesis decreases, etc. Senescent cells are no longer able to replicate, but they remain metabolically active and are positive for senescence-related β-galactosidase activity, which is considered a biomarker of cellular senescence. The Cellular Senescence β-Galactosidase Staining Kit is a kit for staining and detection of senescent cells or tissues based on the up-regulation of the level of senescence-related β-galactosidase activity during senescence. Using X-Gal as the substrate, at pH 6.0, aging-specific β-galactosidase catalyzes the production of dark blue products, so that cells or tissues expressing β-galactosidase turning blue can be observed under light microscope. This kit is suitable for senescence detection of cultured cells and tissue sections, but stains only senescent cells, not presenescent cells, quiescent cells, immortal cells or tumor cells.\u003cbr\u003e\u003cstrong\u003eProduct Components:\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 98.2004%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36.0816%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 28.8992%; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd style=\"width: 72.7484%; text-align: center;\"\u003eStorage conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36.0816%; text-align: center;\"\u003e10 × Fixation Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 28.8992%; text-align: center;\"\u003e15mL\u003c\/td\u003e\n\u003ctd style=\"width: 72.7484%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36.0816%; text-align: center;\"\u003e10 × PBS\u003c\/td\u003e\n\u003ctd style=\"width: 28.8992%; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 72.7484%; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36.0816%; text-align: center;\"\u003eReagent A\u003c\/td\u003e\n\u003ctd style=\"width: 28.8992%; text-align: center;\"\u003e1.5 mL\u003c\/td\u003e\n\u003ctd style=\"width: 72.7484%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36.0816%; text-align: center;\"\u003eReagent B\u003c\/td\u003e\n\u003ctd style=\"width: 28.8992%; text-align: center;\"\u003e1.5 mL\u003c\/td\u003e\n\u003ctd style=\"width: 72.7484%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36.0816%; text-align: center;\"\u003eReagent C\u003c\/td\u003e\n\u003ctd style=\"width: 28.8992%; text-align: center;\"\u003e1.5 mL\u003c\/td\u003e\n\u003ctd style=\"width: 72.7484%; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 36.0816%; text-align: center;\"\u003eX-Gal solution\u003c\/td\u003e\n\u003ctd style=\"width: 28.8992%; text-align: center;\"\u003e6mL\u003c\/td\u003e\n\u003ctd style=\"width: 72.7484%; text-align: center;\"\u003e-20 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cstrong\u003eProduct Features:\u003c\/strong\u003e\u003cbr\u003eSimple, optimized experimental method.\u003cbr\u003eEasy and quick to operate.\u003cbr\u003eIt is suitable for senescence detection of cultured cells and tissue sections.\u003cbr\u003eOnly senescent cells were stained, not presenescent cells, quiescent cells, immortal cells, or tumor cells.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. This kit can only stain senescent cells, not presenescent cells, quiescent cells, immortal cells or tumor cells.\u003cbr\u003e2. X-Gal solution will freeze when stored at-20 °C or 4 °C. It can be completely melted at room temperature or 37 °C water bath for 2-5 minutes and shaken properly.\u003cbr\u003e3. Cell senescence β-galactosidase staining reaction depends on specific pH conditions and cannot be used in CO\u003csub\u003e2\u003c\/sub\u003eThe staining reaction was carried out in an incubator. Higher concentration of CO in incubator\u003csub\u003e2\u003c\/sub\u003eCan affect the pH of the Staining Working Solution and cause dyeing failure. For multi-hole plates, they can be sealed with sealing film or plastic wrap to prevent evaporation.\u003cbr\u003e4. If there is precipitation after the reagent is thawed or before use, it must be ensured that the precipitation is completely dissolved before use. When Reagent C is just removed from the kit, there may be a small amount of precipitate at the bottom of the tube, which is normal. After thorough mixing, the precipitate will completely dissolve, and it must be used after complete dissolution. When preparing the Staining Working Solution, a small amount of flocculent precipitate may also appear, which will be completely dissolved after shaking and mixing, and it must be ensured that it is completely dissolved before use.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eOwn instruments, reagents, consumables: \u003c\/strong\u003e\u003cbr\u003emicroscope \u003cbr\u003eincubator without CO\u003csub\u003e2\u003c\/sub\u003e37℃ \u003cbr\u003epipet and tip, polypropylene tube (15 or 50 mL)\u003cbr\u003eVarious glass utensils for preparation of reagents and buffer solutions \u003cbr\u003edeionized water, glycerol \u003cbr\u003esealing membrane, 6 orifice  p \u0026gt;\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003ereagent preparation: \u003cbr\u003e strong \u0026gt;1 \u0026amp; times; PBS: diluted to 1\u0026amp;times with deionized water before use; PBS, equilibrated to room temperature; Store at 4 ° C. \u003cbr\u003e1× Fixation Buffer: Before use, use 1× Dilute to 1\u0026amp;times with PBS; Fixation Buffer, equilibrated to room temperature; Store at -20 ° C. \u003cbr\u003eReagent A: ready-to-use type; Before use, equilibrate to room temperature; Store at -20 ° C. \u003cbr\u003eReagent B: ready-to-use type; Before use, equilibrate to room temperature; Store at -20 ° C. \u003cbr\u003eReagent C: ready-to-use type; Before use, equilibrate to room temperature; Store at -20 ° C. \u003cbr\u003eX-Gal solution: ready-to-use type; Before use, balance to 37 ° C; Store at -20 ° C away from light. It is very important to heat the X-gal solution at 37 ° C for 1h to avoid the formation of aggregates, which may interfere with the visualization of stained cells. \u003cbr\u003eStaining Working Solution: press 10µ L Reagent A, 10 µ L Reagent B,10 µ L Reagent C ,50 µ L X-gal solution and 920 µ L 1× The reagent was mixed in the ratio of PBS and then adjusted to pH 6.0 with NaOH or HCl. \u003cbr\u003e\u003cstrong\u003eNote: When preparing the staining solution, use a container of polypropylene material or a glass container. A small amount of flocculated precipitate may occur, and it will dissolve completely after shaking and mixing. Make sure it is completely dissolved before use. \u003cbr\u003e\u003c\/strong\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure: \u003cbr\u003eNote: Staining Working Solution is toxic and corrosive to human body. Be careful when handling and pay attention to effective protection. Avoid contact with humans or direct inhalation. \u003c\/strong\u003e\u003cbr\u003eI. For adherent cells: \u003cbr\u003e1, the cells were directly cultured on a cover slip of a 6-well plate and treated as needed. \u003cbr\u003e2, remove the cell culture medium and use 1× The cells were washed twice with PBS. \u003cbr\u003e3, add 1 mL 1\u0026amp;times to each well; The plates were fixed with Fixation Buffer for 15 min at room temperature. For other types of culture plates, the amount of fixation buffer and subsequent solution is operated according to this ratio. \u003cbr\u003e4, remove the cell fixative and use 1× The cells were washed 3 times with PBS. \u003cbr\u003e5, blot out 1× PBS, 1 mL Staining Working Solution was added to each well. \u003cbr\u003e6, the cells were incubated overnight in a 37 ° C incubator in the absence of CO\u003csub\u003e2\u003c\/sub\u003euntil they stained blue. \u003cbr\u003e\u003cstrong\u003eNote: Seal the 6-well plate with a sealing membrane to prevent drying of the cells, and choose the appropriate time for staining. It is not possible to grow cells in an incubator containing CO2 \u003c\/strong\u003ebecause the cell senescence stain is pH dependent. \u003cbr\u003e7, the cells were placed under a light microscope for observation. The number of blue cells and total cells was counted, and the expression of \u0026amp;beta was calculated. -Percentage of cells expressing galactosidase (senescent cells). After Staining, if the Staining Working Solution cannot be observed and counted in time, the staining working solution can be removed and 2 mL of 1\u0026amp;times added; PBS, 4 ° C can be stored for several days or cover cells with 70% glycerol solution at 4 ° C for long-term storage. \u003cbr\u003e\u003cbr\u003e2. For suspended cells: \u003cbr\u003e1, 1000 rpm for 5 min to collect cells into 1.5 mL centrifuge tube, and use 1× PBS  Wash 2 times.   \u003cbr\u003e2, add 1 mL 1\u0026amp;times to each tube; The tubes were fixed with Fixation Buffer for 15 min at room temperature. The cells can be fixed by slowly shaking them on a shaker to avoid clumping. \u003cbr\u003e3, 1000 rpm, after centrifugation for 5 min, remove the cell fixative and use 1× The cells were washed 3 times with PBS. \u003cbr\u003e4, blot out 1× PBS, 1 mL Staining Working Solution was added to each tube. \u003cbr\u003e5, the cells were incubated overnight in a 37 ° C incubator in the absence of CO\u003csub\u003e2\u003c\/sub\u003euntil they stained blue. \u003cbr\u003e6, some of the stained cells were dropped onto a slide or inside a 6-well plate and observed under an ordinary light microscope. The number of blue cells and total cells was counted, and the expression of \u0026amp;beta was calculated. -Percentage of cells expressing galactosidase (senescent cells). If the Staining Working Solution cannot be observed and counted in time, the staining working solution can be removed and 2 mL 1\u0026amp;times added; PBS, 4 ° C can be stored for several days or cover cells with 70% glycerol solution at 4 ° C for long-term storage. \u003c\/p\u003e\n\u003cp\u003eIII. For tissue sections: \u003cbr\u003eFor paraffin sections, deparaffinization and hydration were performed first according to conventional methods. For frozen sections, follow the following steps directly. \u003cbr\u003e1, for prepared tissue sections, add the appropriate volume of 1× Fixation Buffer, appropriate to fully cover the tissue, fixed at room temperature for 15mim\u003cbr\u003e2, with 1× The tissues were soaked in PBS and washed 3 times. \u003cbr\u003e3, absorb for 1× PBS, and the appropriate amount of Staining Working Solution was added. \u003cbr\u003e4, the cells were incubated overnight in a 37 ° C incubator without CO\u003csub\u003e2\u003c\/sub\u003euntil they stained blue. \u003cbr\u003e5, the tissue sections were placed under a light microscope for observation. The number of blue cells and total cells was counted, and the expression of \u0026amp;beta was calculated. -Percentage of cells expressing galactosidase (senescent cells). After staining, if it cannot be observed in time, cover the tissue sections with 70% glycerol solution and store them at 4 ° C for a long time. \u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eStored at-20 ℃, shelf life is 12 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eCell and Tissue Sections\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100T","offer_id":41518140620875,"sku":"abs50063-100T","price":147.0,"currency_code":"USD","in_stock":true}]},{"product_id":"mitochondrial-permeability-transition-pore-assay","title":"Mitochondrial Permeability Transition Pore Assay","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSelf-brought instruments, reagents and consumables:\u003c\/strong\u003e\u003cbr\u003eCell culture plate, adjustable pipette gun and tip\u003cbr\u003eCentrifuge\u003cbr\u003eFluorescence microscope or flow cytometry\u003cbr\u003ePBS\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eReagent Preparation:\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003eCalcein AM staining solution: Add 1 μL of Calcein AM (1000X) per 1 mL of Assay buffer and mix well.\u003cbr\u003e\u003cstrong\u003eNote: The final concentration of Calcein AM needs to be optimized by pre-experiment according to different cell lines and experimental systems. The recommended working concentration of Calcein AM is 1 ×, which can be adjusted between 0.5 ×-5 ×.\u003c\/strong\u003e\u003cbr\u003eFluorescence quenching solution: 10 μL of CoCl per 1 mL of Calcein AM staining solution\u003csub\u003e2\u003c\/sub\u003e(100 ×), mix well.\u003cbr\u003e\u003cstrong\u003eNote: CoCl\u003csub\u003e2\u003c\/sub\u003eThe final concentration is recommended to be 1 ×, and the quenching effect is usually better at this time. CoCl\u003csub\u003e2\u003c\/sub\u003eThe final concentration can also be appropriately optimized according to the type of cells used in the experiment to find the best quenching effect, and can be adjusted between 0.1 ×-1 ×.\u003c\/strong\u003e\u003cbr\u003eIonomycin control: Add 5 μL of Ionomycin (200 ×) per 1 mL of Fluorescence quenching solution and mix well.\u003cbr\u003e\u003cstrong\u003eNote: The final concentration of Ionomycin is recommended to be 1 × and can also be adjusted between 0.5 ×-5 ×.\u003cbr\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure:\u003cbr\u003eNote: This kit (100 T) 96-well plate can detect 1000 T when 100 μL of the detection system per well.\u003cbr\u003e\u003c\/strong\u003e1. Flow cytometry detection:\u003cbr\u003e1. Treat the cells in the expected method;\u003cbr\u003e2. For non-adherent cells, centrifuge at 300 g for 5 min to collect the cells, wash them twice with PBS, and discard the PBS. For adherent cells, cells were first digested with trypsin (without EDTA), then centrifuged at 300 g for 5 min, centrifuged to collect cells, washed twice with PBS, and discarded PBS.\u003cbr\u003e\u003cstrong\u003eNote: Prepare cell samples suspended in Assay Buffer for negative control at flow cytometry.\u003cbr\u003e\u003c\/strong\u003e3. Add appropriate volumes of Calcein AM staining solution, Fluorescence quenching solution, and Ionomycin control to resuspend cells, so that the cell density is about 1 × 10\u003csup\u003e6\u003c\/sup\u003e\/mL, incubate at 37 ℃ in the dark for 30-45 min, and the optimal incubation time of different cells is different.\u003cbr\u003e4. After incubation, centrifuge at 300g for 5 minutes to collect the cells. Add 1 mL of Assay Buffer to each sample, gently resuspend, and centrifuge at 300 g for 5 min to collect cells.\u003cbr\u003e5. Cells were resuspended with 400 μL Assay Buffer and analyzed by flow cytometry.\u003c\/p\u003e\n\u003cp\u003e2. Fluorescence microscope detection:\u003cbr\u003e1. For adherent cells:\u003cbr\u003e(1) Cultivate the cells on a suitable well plate in a CO2 cell incubator at 37 °C for at least 24 h, and then perform subsequent experiments.\u003cbr\u003e(2) Treat the cells with the expected method, and incubate the cells without inducer to establish a negative control group.\u003cbr\u003e(3) The cells were washed twice with PBS.\u003cbr\u003e(4) Add appropriate volumes of Calcein AM staining solution, Fluorescence quenching solution, and Ionomycin control to the cells, usually 100 μL per well for 96-well plates, 250 μL per well for 24-well plates, 500 μL per well for 12-well plates, 1 mL per well for 6-well plates, and incubate at 37 ℃ in the dark for 30-45 minutes. The optimal incubation time of different cells is different.\u003cbr\u003e(5) After incubation, replace it with fresh preheated culture medium at 37 ℃, and incubate at 37 ℃ for 30 minutes in the dark to ensure that the cellular esterase fully hydrolyzes Calcein AM to generate green fluorescent Calcein.\u003cbr\u003e(6) Wash with PBS 2-3 times, and then add Assay Buffer to observe under a fluorescence microscope (Calcein AM is green fluorescence, Ex\/Em = 494\/517 nm).\u003c\/p\u003e\n\u003cp\u003e2. For suspension cells:\u003cbr\u003e(1) Cells were processed in the expected manner and counted.\u003cbr\u003e(2) Take 300 g of appropriate cells and centrifuge them for 5 min, discard the supernatant, wash the cells twice with PBS, and discard the PBS.\u003cbr\u003e(3) Resuspend cells with appropriate volumes of Calcein AM staining solution, Fluorescence quenching solution and Ionomycin control, respectively, so that the cell density is about 1 × 10\u003csup\u003e6\u003c\/sup\u003e\/mL, incubate at 37 ℃ in the dark for 30-45 min, and the optimal incubation time of different cells is different.\u003cbr\u003e(4) Centrifuge at 300g for 5 minutes, aspirate the supernatant, slowly add 1mL of preheated culture medium at 37 °C to resuspend the cells, and incubate at 37 °C for 30 minutes in the dark to ensure that the intracellular esterase fully hydrolyzes Calcein AM to generate green fluorescent Calcein.\u003cbr\u003e(5) Centrifuge at 300g for 5 minutes, aspirate most of the culture medium, resuspend the cells and smear them, and observe under a fluorescence microscope.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e \u003c\/strong\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003emitochondrial permeablity transition pore (MPTP), also known as mitochondrial giant channel (magachannel), is a non-selective highly conductive channel existing between the inner and outer membranes of mitochondria, composed of a variety of protein complexes. Mitochondrial permeability transition pores (MPTP) may be involved in the release of mitochondrial components during cell death. The mitochondrial inner membrane of normal cells can maintain normal mitochondrial potential gradient to ensure cell respiration and energy supply. With Ca\u003csup\u003e2+\u003c\/sup\u003eThe intake and release of, a low-conductivity osmotic switching hole switches back and forth between opening and closing. When cells undergo apoptosis and pathological death, the mitochondrial membrane potential switching pore permeability is changed, and Ca\u003csup\u003e2+\u003c\/sup\u003eOverload, oxidation of mitochondrial glutathione, increase of reactive oxygen levels, including subsequent release of cytochrome C, and decrease of mitochondrial membrane potential will all lead to the activation of mitochondrial permeability transition pores. Mitochondrial membrane permeability transition pore assay kit is a more direct detection method for mitochondrial permeability transition pore openness than only mitochondrial membrane potential analysis. The principle is: first, Calcein AM is loaded through passive transportation. The latter is a cell staining reagent that fluorescently labels living cells. It can easily penetrate the living cell membrane and be sheared by esterases in the cell to form a membrane-non-permeable polar molecule Calcein, which is retained in the cell, causing the cytoplasm, including mitochondria, to emit strong green fluorescence. Add CoCl\u003csub\u003e2\u003c\/sub\u003eAfter that, the fluorescence from the cytoplasm was reduced by CoCl\u003csub\u003e2\u003c\/sub\u003eQuenching, leaving only fluorescence within the mitochondria. As controls, cells can be loaded with Calcein AM and CoCl\u003csub\u003e2\u003c\/sub\u003eAt the same time, it was treated with Ionomycin to load the cells with more Ca\u003csup\u003e2+\u003c\/sup\u003e, causing activation of mitochondrial permeability transition pores to quench mitochondrial fluorescence.\u003cbr\u003eProduct Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 68.021%; height: 110px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 37.3995%; height: 22px;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 16.1991%; height: 22px;\"\u003e50T\u003c\/td\u003e\n\u003ctd style=\"width: 18.2248%; height: 22px;\"\u003e100T\u003c\/td\u003e\n\u003ctd style=\"width: 21.0774%; height: 22px;\"\u003eStorage conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 37.3995%; height: 22px;\"\u003eCalcein AM (1000 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 16.1991%; height: 22px;\"\u003e50 μL\u003c\/td\u003e\n\u003ctd style=\"width: 18.2248%; height: 22px;\"\u003e100 μL\u003c\/td\u003e\n\u003ctd style=\"width: 21.0774%; height: 22px;\"\u003e-20 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 37.3995%; height: 22px;\"\u003eCoCl\u003csub\u003e2\u003c\/sub\u003e（100×）\u003c\/td\u003e\n\u003ctd style=\"width: 16.1991%; height: 22px;\"\u003e0.5 mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.2248%; height: 22px;\"\u003e1mL\u003c\/td\u003e\n\u003ctd style=\"width: 21.0774%; height: 22px;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 37.3995%; height: 22px;\"\u003eIonomycin (200 ×)\u003c\/td\u003e\n\u003ctd style=\"width: 16.1991%; height: 22px;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 18.2248%; height: 22px;\"\u003e500 μL\u003c\/td\u003e\n\u003ctd style=\"width: 21.0774%; height: 22px;\"\u003e-20 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 37.3995%; height: 22px;\"\u003eAssay Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 16.1991%; height: 22px;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 18.2248%; height: 22px;\"\u003e100 mL × 2\u003c\/td\u003e\n\u003ctd style=\"width: 21.0774%; height: 22px;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003eProduct Features:\u003cbr\u003eHigh safety: almost no toxicity to cells;\u003cbr\u003eCompatibility with multiple detection methods: Can be applied to flow cytometers and fluorescence microscopes.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTechnical Index\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eFluorescence excitation\/emission: Calcein-AM: Ex\/Em = 494\/517 nm\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e\u003cstrong\u003eshould bring along their own instruments, reagents, consumables:  strong \u0026gt;\u003cbr\u003ecell culture plate, adjustable moving fluid and spear gun \u003cbr\u003e\u003cbr\u003ethe centrifuge fluorescence microscope and flow cytometry instrument \u003cbr\u003ePBS  p \u0026gt;\u003cp\u003e\u003cstrong\u003ereagent preparation:  strong \u0026gt; p \u0026gt;\u003cp\u003eCalcein AM staining solution: each 1 mL Assay buffer with 1 \u0026amp; mu; L of Calcein AM (1000X)  And blending. \u003cbr\u003e\u003cstrong\u003enote: Calcein AM final concentration should be based on different cell lines and the experimental system is optimized through an experiment. Calcein AM  The recommended working concentration of calcein is 1× , can be at 0.5× -5× Make adjustments between. \u003c\/strong\u003e\u003cbr\u003eFluorescence quenching solution: 10 \u0026amp;mu per 1 mL Calcein AM staining solution; L CoCl\u003csub\u003e2\u003c\/sub\u003e(100×) And blending. \u003cbr\u003e\u003cstrong\u003enote: CoCl \u003csub\u003e2  sub \u0026gt;recommended the final concentration of 1 \u0026amp; times; In general, the quenching effect is better at this time. The final concentration of CoCl\u003csub\u003e2 \u003c\/sub\u003ecan also be appropriately optimized according to the type of cells used in the experiment to find the best quenching effect, which can be in 0.1× -1× Make adjustments between. \u003c\/sub\u003e\u003c\/strong\u003e\u003cbr\u003eIonomycin control: Add 5\u0026amp;mu to every 1 mL Fluorescence quenching solution; L of   Ionomycin (200×) , mix well. \u003cbr\u003e\u003cstrong\u003eNote: The recommended final concentration of Ionomycin is 1× , can also be used at 0.5× -5× Make adjustments between. \u003cbr\u003e\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental procedure: \u003cbr\u003eNote: The detection system of this kit (100 T) 96-well plate is 100 μ It can detect 1000 T at L. \u003cbr\u003e\u003c\/strong\u003eI. Flow cytometry: \u003cbr\u003e1. The cells were treated as expected; \u003cbr\u003e2, for non-adherent cells, cells were collected by centrifugation at 300 g for 5 min, washed twice with PBS, and the PBS was discarded. For adherent cells, cells were first digested with trypsin (without EDTA), then centrifuged at 300 g for 5 min, collected by centrifugation, washed twice with PBS, and discarded with PBS. \u003cbr\u003e\u003cstrong\u003eNote: A sample of cells suspended in Assay Buffer should be prepared to be used as a negative control during flow cytometry. \u003cbr\u003e\u003c\/strong\u003e3. Add the appropriate volume of Calcein AM staining solution, Fluorescence quenching solution and Ionomycin control to resuspend the cells, respectively. The cell density was about 1× 10 \u003csup\u003e6  sup \u0026gt;\/ mL, 37 ℃ avoid light incubation 30-45 min, cells of different optimal incubation time is different. \u003cbr\u003e4, After completion of incubation, cells were collected by centrifugation at 300g for 5 min. 1mL Assay Buffer was added to each sample, gently resuspended, and cells were collected by centrifugation at 300g for 5 min. \u003cbr\u003e5, using 400 μ L Assay Buffer after heavy suspension cells, cells in the analysis.  p \u0026gt;\u003cp\u003e2, fluorescence microscope detection: \u003cbr\u003e1, the adherent cells: \u003cbr\u003e(1) on the suitable orifice plate culture cell, cell cultivation in 37 ℃ in CO2 at least 24 h, then follow-up experiments. \u003cbr\u003e(2) Cells were treated with the expected methods and incubated without inducers to establish a negative control group. \u003cbr\u003e(3) Cells were washed twice with PBS. \u003cbr\u003e(4) Add appropriate volumes of Calcein AM staining solution, Fluorescence quenching solution, and Ionomycin control to the cells, respectively. Usually 100\u0026amp;mu was added to each well of 96-well plate; L, add 250 \u0026amp;mu to each well of a 24-well plate; L, add 500 \u0026amp;mu to each well of 12-well plate; 1 mL was added to each well of L, 6-well plates and incubated at 37 ° C in the dark for 30-45 min, with the optimal incubation time for different cells varying. \u003cbr\u003e(5) At the end of the incubation, the cells were replaced with fresh culture medium preheated at 37℃ and incubated at 37℃ in the dark for another 30min to ensure that the intracellular esterase could fully hydrolyze Calcein AM to produce Calcein with green fluorescence. \u003cbr\u003e(6) Wash with PBS for 2-3 times, then add Assay Buffer and observe under fluorescence microscope (Calcein AM is green fluorescence, Ex\/Em=494\/517nm). \u003c\/p\u003e\n\u003cp\u003e2, for suspended cells: \u003cbr\u003e(1) Cells were treated as expected and counted. \u003cbr\u003e(2) Appropriate cells were centrifuged at 300 g for 5 min, the supernatant was discarded, and \u0026amp;nbsp was used; The cells were washed twice with PBS and the PBS was discarded. \u003cbr\u003e(3) The cells were resuspended by adding appropriate volumes of Calcein AM staining solution, Fluorescence quenching solution, and Ionomycin control, respectively, so that the cell density was about 1× 10\u003csup\u003e6\u003c\/sup\u003e\/mL, and the cells were incubated at 37 ° C in the dark for 30-45 min. The optimal incubation time was different for different cells. \u003cbr\u003e(4) The cells were centrifuged at 300g for 5 min, the supernatant was removed, and the cells were resuspended by slowly adding 1mL of preheated culture medium at 37℃. The cells were incubated at 37℃ for another 30min in the dark to ensure that the intracellular esterase could fully hydrolyze Calcein AM to produce Calcein with green fluorescence. \u003cbr\u003e(5) The cells were centrifuged at 300g for 5 min, most of the culture medium was removed by suction, and the cells were resuspended for smear and observed under a fluorescence microscope. \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e  \u003c\/strong\u003e\u003c\/p\u003e\u003c\/sup\u003e\u003c\/p\u003e\u003c\/strong\u003e\u003c\/p\u003e\u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e-20 ℃, protected from light, shelf life for 12 months.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50T","offer_id":41518140850251,"sku":"abs50064-50T","price":152.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41518140883019,"sku":"abs50064-100T","price":270.0,"currency_code":"USD","in_stock":true}]},{"product_id":"celltiter-turbo-2-0-luminescentcell-viability-assay","title":"CTG-L™ 2D Luminescent Cell Viability Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e\u003cstrong\u003e1. Reagent preparation\u003c\/strong\u003e\u003cstrong\u003e：\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eReagent melt:\u003c\/strong\u003eThe reagents were removed the day before the experiment and left at 4 °C overnight to melt. The reagents can also be taken out on the day of the experiment and melted at room temperature, or melted in a 22 °C water bath, but\u003cstrong\u003eIt is necessary to note that the water temperature should not exceed\u003c\/strong\u003e\u003cstrong\u003e25\u003c\/strong\u003e\u003cstrong\u003e℃\u003c\/strong\u003e。\u003cbr\u003e\u003cstrong\u003eEquilibrate to room temperature:\u003c\/strong\u003eIf the reagent melts at non-room temperature, it can be placed in a 22 ℃ water bath before use to ensure that the reagent is equilibrated to room temperature before being used for testing.\u003cbr\u003eNote: Generally, it takes about 10 minutes for 5mL packaging; The 50 mL packaging takes about 20 minutes.\u003cbr\u003e\u003cstrong\u003eMix well:\u003c\/strong\u003eBefore use\u003cstrong\u003eGentle upside down\u003c\/strong\u003e\u003cstrong\u003e5\u003c\/strong\u003e\u003cstrong\u003eTime\u003c\/strong\u003eThe solution was mixed evenly.\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003cstrong\u003e2. Detection steps:\u003cbr\u003e\u003c\/strong\u003e1. Cell culture: Use a 96-well plate suitable for chemiluminescence detection, and inoculate 100μL cells per well (determine the initial cell density according to the culture time, and the number of cells per well should not exceed 100,000 during detection). At the same time, set a well of culture medium containing no cells as a negative control. The concentration gradient of the cells can also be set to obtain the best experimental results. 37 °C, 5% CO\u003csub\u003e2\u003c\/sub\u003eCulture the cells and treat the cells with dosing at the appropriate time as needed.\u003cbr\u003e2. (Optional) Preparation of ATP standard curve:\u003cbr\u003eThe prepared ATP standard solution was diluted with PBS to the appropriate concentration gradient, and 100 μL of the standard was added to each well of the 96-well plate.\u003cbr\u003e3. Cell viability detection\u003cbr\u003e(1) Thaw the frozen luminescence detection reagent, equilibrate it to room temperature (or 22 ℃ constant temperature water bath equilibrium), and take out the volume of detection reagent used for the current experiment\u003cbr\u003e(2) Take out the cell culture plate and balance it at room temperature for 10 minutes (or balance it in a constant temperature water bath at 22 ℃, the time should not be too long, try to control it within 30 minutes)\u003cbr\u003e(3) Add 100μL of detection reagent to each well of 96-well plate (due to the edge effect of the well, the luminescence signal may be unstable, so it is not recommended to plate at the edge)\u003cbr\u003e(4) Shake at room temperature for 2 minutes to promote cell lysis;\u003cbr\u003e(5) Place at room temperature for 10 minutes to stabilize the luminescent signal;\u003cbr\u003e(6) Use a multifunctional microplate reader for chemiluminescence detection. Set the corresponding parameters according to the requirements of the instrument. The detection time of each hole is generally 0.25-1s. The specific adjustments need to be made according to the detection sensitivity of the instrument;\u003cbr\u003e(7) Calculate the relative viability of cells according to the chemiluminescence reading, or calculate the ATP content according to the ATP standard curve to obtain the relative viability of cells.\u003cbr\u003eNote: The detection effect varies with different types of cells. For some cells with particularly high ATP content, the chemiluminescence reading may continue to increase when the number of cells reaches more than 100,000, but the linear relationship is lost.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eATP cell viability detection kit; 2D Cell Viability Detection Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e2D Luminescent Cell Viability Assay uses ATP-dependent luciferase-catalyzed luciferin luminescence reaction to measure intracellular ATP content through chemiluminescence signal, so as to detect cell viability or quantitatively detect the number of viable cells. The detection has wide linear range, high sensitivity and good stability. In 96-well plates, there is a good linear relationship in the range of 100 to 100,000 cells, but the upper limit of the number of detected cells will be different. In addition, the operation is simple, the detection reagents provided in the kit are ready-to-use, with stable readings and fast detection speed. It only takes about 10 minutes to complete the detection, and there is no need to wash the cells, and there is no need to replace or remove the culture medium. Compared with other common cell viability measurement methods, such as Calcein-AT, CCK-8, etc., 2D Luminescent Cell Viability Assay is simpler and faster.  \u003cbr\u003eThe 2D Luminescent Cell Viability Assay has high detection sensitivity and wide linear range, and is compatible with small sample detection and high-throughput screening of large samples.  \u003cbr\u003e1. Convenient and fast: The detection reagents provided in the kit are ready-to-use, with stable readings and fast detection speed. It only takes about 10 minutes to complete the detection;  \u003cbr\u003e2. High sensitivity: can detect ATP with a minimum of 10nmol;  \u003cbr\u003e3. Wide linear range: In the 96-well plate, there is a good linear relationship in the range of 100 to 100,000 cells, but the upper limit of the number of detected cells will be different;  \u003cbr\u003e4. High throughput: It is compatible with the detection of a small amount of samples and the high throughput screening of a large number of samples.\u003cdiv\u003e\u003cimg style=\"display: block; margin-left: auto; margin-right: auto;\" src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240312\/1e959b9d92ce44879f45c6ec48822124.png\" alt=\"\" width=\"571\" height=\"180\"\u003e\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e\u003cstrong\u003e1, temperature:\u003c\/strong\u003eThe reagent contains luciferase, and repeated freezing and thawing will affect its activity. It is recommended to store at-20 ℃ in the dark from light after dispensing. Reagents and cell samples should be balanced to room temperature before use to avoid the influence of enzyme catalytic effect;\u003cbr\u003e\u003cstrong\u003e2. Chemical factors:\u003c\/strong\u003eThe reaction rate and luminescence intensity of luciferase are affected by the chemical environment. There are differences in luminescence intensity and attenuation rate among different types of media and serum. In addition, high drug content may interfere with the luciferase reaction, thus affecting the luminescence signal. It is recommended to set up cell culture medium control wells containing drug to eliminate solvent interference. If the culture medium has a great influence on the luminescence intensity, it can be removed before detection and washed once with PBS;\u003cbr\u003e\u003cstrong\u003e3. Light sensitivity:\u003c\/strong\u003eThis reagent is sensitive to light and will accelerate the attenuation of luminous intensity if exposed to light during storage. If the reagent is transferred from the original container, make sure to keep it protected from light;\u003cbr\u003e\u003cstrong\u003e4. ATP\u003c\/strong\u003e\u003cstrong\u003eContamination:\u003c\/strong\u003eIt is recommended to wear a mask and latex gloves during operation and avoid contact with surfaces and equipment that may be contaminated. Avoid inserting the tip of the gun tip into the vial multiple times during operation.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e\u003cstrong\u003eI. Reagent preparation \u003c\/strong\u003e\u003cstrong\u003e: \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eReagent melting: \u003c\/strong\u003eThe day before the experiment, the reagent was removed and placed at 4 ° C overnight to melt. The reagent can also be removed on the day of the experiment and melted at room temperature, or placed in a 22℃ water bath to melt, but \u003cstrong\u003eneed to note that the water temperature should not exceed \u003c\/strong\u003e\u003cstrong\u003e25\u003c\/strong\u003e\u003cstrong\u003e℃\u003c\/strong\u003e. \u003cbr\u003e\u003cstrong\u003eequilibrate to room temperature: \u003c\/strong\u003eIf the reagent melts under non-room temperature conditions, it can be placed in a 22℃ water bath before use to ensure that the reagent equilibrates to room temperature before being used for detection. \u003cbr\u003eNote: Generally, it takes about 10 minutes for 5mL packaging; 50mL packaging takes about 20min. \u003cbr\u003e\u003cstrong\u003eMix: \u003cstrong\u003egently invert \u003c\/strong\u003e\u003cstrong\u003e5 \u003c\/strong\u003e\u003cstrong\u003etimes \u003c\/strong\u003eto mix the solution well before use. \u003cbr\u003e\u003cstrong\u003eII. Detection steps: \u003cbr\u003e\u003c\/strong\u003e1. Cell culture: use a 96-well plate suitable for chemiluminescence detection, and inoculate 100\u0026amp;mu in each well; L cells (according to the culture time to determine the initial seeding cell density, the number of cells per well should not exceed 100 000), at the same time, set up the culture medium without cells as a negative control. The concentration gradient of cells can also be set to obtain the best experimental results. Cells were cultured at 37 ° C with 5% CO\u003csub\u003e2\u003c\/sub\u003eand treated with drugs at the appropriate time as needed. \u003cbr\u003e2. (Optional) Preparation of ATP standard curve: \u003cbr\u003eThe prepared ATP standard solution was diluted into an appropriate concentration gradient with PBS, and 100\u0026amp;mu was added to each well of a 96-well plate; L of the standard. \u003cbr\u003e3, cell viability detection \u003cbr\u003e(1) Thaw and thaw the luminescence method detection reagent, and balance to room temperature (or 22℃ constant temperature water bath equilibrium) \u003cbr\u003e(2) Remove the cell culture plate, room temperature equilibrium for 10 minutes (or 22℃ constant temperature water bath equilibrium, the time should not be too long, Try to control within 30 minutes) \u003cbr\u003e(3) Add 100\u0026amp;mu to each well of 96-well plate; L detection reagent (due to the edge effect of the well, it may cause the luminescence signal to be unstable, and it is not recommended to spread the plate at the edge) \u003cbr\u003e(4) shake at room temperature for 2 minutes to promote the lysis of cells; \u003cbr\u003e(5) place at room temperature for 10 minutes to make the luminescence signal tend to be stable; \u003cbr\u003e(6) chemiluminescence detection was performed using a multifunctional microplate reader. Set the corresponding parameters according to the requirements of the instrument. The detection time of each well is generally 0.25-1s, and the specific need to be adjusted appropriately according to the detection sensitivity of the instrument. \u003cbr\u003e(7) Relative cell viability was calculated according to the chemiluminescence reading, or ATP content was calculated according to the ATP standard curve to obtain the relative cell viability. \u003cbr\u003eNote: The detection effect varies with the type of cells. For some cells with particularly high ATP content, the chemiluminescence reading may continue to increase when the number of cells reaches 100,000, but the linear relationship may be lost. \u003c\/strong\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eStore at-20 °C protected from light. In order to ensure the best performance, it is recommended to place it at-70 ℃ for long-term storage.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eIt is suitable for activity detection of bioactive factors, large-scale anti-tumor drug screening, cell proliferation, cytotoxicity test, etc.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50mL×2","offer_id":43212776603723,"sku":"abs50065-50mL×2","price":159.0,"currency_code":"USD","in_stock":true},{"title":"10mL","offer_id":41820546269259,"sku":"abs50065-10mL","price":40.0,"currency_code":"USD","in_stock":true},{"title":"100mL","offer_id":41820546302027,"sku":"abs50065-100mL","price":159.0,"currency_code":"USD","in_stock":true},{"title":"100mL×10","offer_id":43212776636491,"sku":"abs50065-100mL×10","price":1200.0,"currency_code":"USD","in_stock":true}]},{"product_id":"ao-pi-double-staining","title":"AO\/PI Double Staining","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Reagent preparation: \u003cbr\u003e(1) PBS buffer (pH7.4) \u003cbr\u003e(2) or HbSS (hank's balanced salt solution) \u003cbr\u003e2. Consumables preparation \u003cbr\u003e(1) centrifuge tube \u003cbr\u003e(2) suction tip \u003cbr\u003e(3) disposable gloves \u003cbr\u003e3. Dyeing working solution preparation: \u003cbr\u003e(1) according to the number of samples, prepare the staining buffer according to the following ratio. Take 100μ L reagent C used 900μ Dilute with L pure water and mix well to form staining buffer\u003cbr\u003e(2) every 500 mu; L staining buffer, add 5ul Ao staining solution and 10ul PI staining solution, and mix well to form the staining working solution\u003cbr\u003e4. Suspension cell staining \u003cbr\u003e(1) collect sample cells, and the number of cells is within 10x105\u003cbr\u003e(2) wash the cells twice with PBS\u003cbr\u003e(3) use 500 mu; L staining working solution to resuspend the cells\u003cbr\u003e(4) incubate for 10-20 minutes at 4 ℃ in the dark after gently mixing\u003cbr\u003e(5) wash the cells with PBS\u003cbr\u003e(6) the results were detected by fluorescence microscope or flow cytometry\u003cbr\u003e5. In situ staining of adherent cells\u003cbr\u003e(1) wash the cells twice with PBS\u003cbr\u003e(2) add an appropriate volume of staining working solution to the cell culture plate or cell climbing piece\u003cbr\u003e(3) incubate the cells at 37 ℃ for 10-20 minutes, and the optimal culture time is different for different cells. 20 min can be used as the initial incubation time, after which the system can be optimized to obtain uniform labeling results\u003cbr\u003e(4) blot the staining working solution, wash the culture plate or coverslip with medium for 2-3 times, cover all cells with preheated medium each time, and then blot the medium\u003cbr\u003e\u003cbr\u003eResult Analysis: \u003cbr\u003eUnder the fluorescence microscope, 488nm excitation light was used for microscopic examination: \u003cbr\u003ethe DNA of normal cells stained with AO was uniform yellow or yellow green, and the morphology and structure were normal\u003cbr\u003ewhen cells are apoptotic, chromatin condenses, and nuclei are fragmented into punctate, which are stained in a dense and dense manner with different sizes\u003cbr\u003eNecrotic cells showed strong red fluorescence\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e  AO\/PI double staining apoptosis detection kit is a commonly used staining method for apoptosis morphology research, which uses AO\/PI probe to double stain the nucleus to detect the state of apoptotic cells\u003cbr\u003e  Acridine orange (AO) is a fluorescent dye with cell membrane permeability. The dye can penetrate the living cell membrane and stain nuclear DNA and RNA. It is different from the binding amount of DNA and RNA in cells. The complex can emit different colors of fluorescence. Ao emits green fluorescence when it is combined with dsDNA, and red fluorescence when it is combined with ssDNA and RNA. When bound to DNA, it is very similar to fluorescein in spectrum, with an excitation maximum of 502nm and an emission maximum of 525nm (green). When it binds to RNA, the excitation maximum shifts to 460nm (blue) and the emission maximum shifts to 650nm (red)\u003cbr\u003e  Under the fluorescence microscope, acridine orange can penetrate the normal cell membrane, making the nucleus green or yellowish green uniform fluorescence; In apoptotic cells, apoptotic bodies are formed due to chromatin pyknosis or fragmentation into fragments of different sizes. Acridine orange causes it to be stained with dense yellow green fluorescence, or yellow green fragment particles; However, the yellow fluorescence of necrotic cells decreased or even disappeared\u003cbr\u003e  Propidium iodide is a DNA binding dye. Its excitation and emission wavelengths are 488nm and 630nm, respectively. It produces red fluorescence, but it has no membrane permeability, cannot penetrate the membrane of living cells, and can only dye dead cells. Therefore, under the fluorescence microscope, normal cells cannot be colored, early apoptotic cells show weak red light, late apoptotic cells show enhanced red light, and necrotic cells show strong red fluorescence.\u003c\/p\u003eProduct components: \u003cbr\u003e\u003ctable style=\"border collapse: collapse; width: 71.0907%; margin: 0 Auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 19.5605%; text align: Center;\"\u003eComponent\u003c\/td\u003e\n\u003ctd style=\"width: 24.4891%; text align: Center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 23.5732%; text align: Center;\"\u003eSpecification\u003c\/td\u003e\n\u003ctd style=\"width: 25.2595%; text align: Center;\"\u003eSave\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 19.5605%; text align: Center;\"\u003eComponent A\u003c\/td\u003e\n\u003ctd style=\"width: 24.4891%; text align: Center;\"\u003e\u003cp\u003eAO staining solution\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 23.5732%; text align: Center;\"\u003e500ul\u003c\/td\u003e\n\u003ctd style=\"width: 25.2595%; text align: Center;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 19.5605%; text align: Center;\"\u003e\u003cp\u003eComponent B\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 24.4891%; text align: Center;\"\u003e\u003cp\u003ePI staining solution\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 23.5732%; text align: Center;\"\u003e1000ul\u003c\/td\u003e\n\u003ctd style=\"width: 25.2595%; text align: Center;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 19.5605%; text align: Center;\"\u003e\u003cp\u003eComponent C\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 24.4891%; text align: Center;\"\u003eReagent C\u003c\/td\u003e\n\u003ctd style=\"width: 23.5732%; text align: Center;\"\u003e10ml\u003c\/td\u003e\n\u003ctd style=\"width: 25.2595%; text align: Center;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. The reagent in the screw cap micro reagent tube should be centrifuged briefly before opening the cap, and the liquid on the inner wall of the cap should be collected to the bottom of the tube to avoid liquid spilling when opening the cap\u003cbr\u003e2. Careful operation is required for cell treatment to avoid artificial damage to cells\u003cbr\u003e3. This staining kit can be used for cells, cell smears, etc. The following is an example of fluorescence microscopy detection of suspension cultured cells. For other methods, please refer to relevant materials\u003cbr\u003e4. Adherent cells can be digested and stained, or directly stained in situ. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e2-8 ℃ protected from light, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplications\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eFlow cytometry; Laser confocal; fluorescence microscope\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100T","offer_id":41518203240523,"sku":"abs9727-100T","price":100.0,"currency_code":"USD","in_stock":true}]},{"product_id":"ao-be-double-staining-kit","title":"AO\/EB Double Staining Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eSelf prepared material:\u003cbr\u003eFluorescence microscope; PBS; Cell counting board; Glass slide; Cover glass slides\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e1. Preparation of staining working solution\u003cbr\u003e\u003c\/strong\u003eBefore the formal experiment, take acridine orange staining solution (reagent A), ethidium bromide staining solution (reagent B), and dilution buffer solution (reagent C), and dilute them to the required AO\/EB staining working solution in the ratio of A: B: C=1:1:8\u003cbr\u003e\u003cstrong\u003e2. Cell preparation\u003cbr\u003e\u003c\/strong\u003eAdherent cells: Digest according to conventional methods. Centrifuge at 1000 rpm for 5 minutes, collect cell precipitates, and wash once with PBS6pieces\/ml\u003cbr\u003e\u003cstrong\u003e3. Staining\u003cbr\u003e\u003c\/strong\u003eEvery 25μ Add 2\u0026amp;mu to the cell suspension; Freshly prepared AO\/EB staining working solution, gently mix evenly. Incubate at room temperature for 5-10 minutes\u003cbr\u003e\u003cstrong\u003e4. Observation\u003cbr\u003e\u003c\/strong\u003eTake a clean slide and add 5-10\u0026amp;mu dropwise; Cover the stained cell suspension with a cover glass. Observe and count using a fluorescent microscope's luciferin filter and a 60x magnification objective. This measurement should be repeated at least three times, with at least 100 total cells observed each time\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e  Acridine orange (AO) is a heterochromatic nucleic acid binding dye with cell membrane permeability. It binds to DNA or RNA by insertion or electrostatic attraction. When combined with double stranded DNA (dsDNA), it emits green fluorescence (ex\/em=502\/525nm), and when combined with single stranded DNA (ssDNA) or RNA, it emits red fluorescence (ex\/em=460\/650nm), with a small amount of binding showing orange red fluorescence. Ethidium bromide (EB) is a polycyclic aromatic hydrocarbon fluorescent small molecule and nucleic acid insertion agent, which is chimeric into the base pairs of double stranded DNA or RNA. It has no base specificity and shows red fluorescence ((ex\/em=518\/605nm). EB is not cell membrane permeable\u003cbr\u003e  Acridine orange (AO) and ethidium bromide (EB) are often used in combination to observe the changes of nucleus and the formation of apoptotic bodies, which are the characteristics of apoptosis. The combination of the two can distinguish living cells, apoptotic cells and necrotic cells. The working principle is that acridine orange can stain both live and dead cells, while EB can only stain cells that have lost membrane integrity. The living cells are uniformly green. Early apoptotic cells are green, and bright green dots are seen in the nuclear energy of cells due to chromosome condensation and nuclear breakage. Late apoptotic cells will also be stained with EB and thus become orange red, but compared with necrotic cells, late apoptotic cells will have concentrated and often broken nuclei. Necrotic cells are also stained orange red, but will have a nuclear morphology similar to that of living cells, without condensed chromosomes.\u003cbr\u003e\u003cbr\u003eProduct Components:\u003c\/p\u003e\n\u003ctable style=\"border collapse: collapse; width: 80.5913%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 14.4655%;\"\u003eComponent\u003c\/td\u003e\n\u003ctd style=\"width: 39.1234%;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 15.7955%;\"\u003eSpecification\u003c\/td\u003e\n\u003ctd style=\"width: 23.6104%;\"\u003eSave\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 14.4655%;\"\u003eComponent A\u003c\/td\u003e\n\u003ctd style=\"width: 39.1234%;\"\u003eAo stain solution acridine orange dye solution\u003c\/td\u003e\n\u003ctd style=\"width: 15.7955%;\"\u003e200ul\u003c\/td\u003e\n\u003ctd style=\"width: 23.6104%;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 14.4655%;\"\u003eComponent B\u003c\/td\u003e\n\u003ctd style=\"width: 39.1234%;\"\u003e\u003cp\u003eEB stain solution ethidium bromide dye solution\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.7955%;\"\u003e200ul\u003c\/td\u003e\n\u003ctd style=\"width: 23.6104%;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 14.4655%;\"\u003eComponent C\u003c\/td\u003e\n\u003ctd style=\"width: 39.1234%;\"\u003e\u003cp\u003eDilution buffer dilution buffer\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.7955%;\"\u003e50ml\u003c\/td\u003e\n\u003ctd style=\"width: 23.6104%;\"\u003e2-8 ℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003e1. It is also commonly used to double stain cells with H O echst 33342, which can produce blue fluorescence after binding to dn a through the living cell membrane, and propidiu m iodide, which can only produce red fluorescence after binding to dn a through dead cells\u003cbr\u003e2. If there is a low-temperature centrifuge for centrifugation, the effect is better\u003cbr\u003e3. During operation, pay attention to reducing the exposure time of reagent (a) and reagent (b) to strong light\u003cbr\u003e4. EB solution has certain toxicity, please be careful\u003cbr\u003e5. For your safety and health, please wear lab clothes and disposable gloves\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e2-8 ℃ protected from light, 6months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100T","offer_id":41518203830347,"sku":"abs9728-100T","price":100.0,"currency_code":"USD","in_stock":true}]},{"product_id":"annexin-v-apc-dapi-apoptosis-kit","title":"Annexin V-APC\/DAPI Apoptosis Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eExperimental procedure:\u003cbr\u003e1. Annexin V-APC and cell incubation\u003cbr\u003e1. Induce apoptosis according to the experimental protocol. Centrifuge at 300 g for 5 min, discard supernatant, collect cells, gently resuspend cells with PBS and count.\u003cbr\u003e2. Centrifugation collection 1-5 × 10\u003csup\u003e5\u003c\/sup\u003eThe supernatant was discarded. The cells were washed once with PBS and the supernatant was discarded after centrifugation.\u003cbr\u003e3. Resuspend the cells with 500 µ L of 1X Binding Buffer.\u003cbr\u003e4. Add 5 µ L Annexin V-APC and 5 µ L DAPI.\u003cbr\u003e5. Gently vortex and mix well, and incubate at room temperature in the dark for 15 minutes.\u003cbr\u003e6. Test immediately. If it cannot be detected in time, please put it on ice to protect from light and complete the test within 1 hour.\u003cbr\u003e2. Flow cytometry quantification\u003cbr\u003eFlow cytometry checks the available APC channel for Annexin V-APC, and the DAPI channel is preferred for DAPI detection, followed by the PacificBlue channel. For adherent cells, the cell culture medium was first collected, the cells were washed once with PBS, and the cells were digested with trypsin, then the collected cell culture medium was added, the cells were gently blown down, the supernatant was discarded by centrifugation, and the cells were resuspended with PBS and counted, and then step 1 2-5 was performed.\u003cbr\u003e3. Fluorescence microscope detection The cell suspension in step 1.5 is coated on a glass slide. Cover cells with a coverslip. If adherent cells are analyzed, the cells are cultured directly on the cell slide. After incubation (step 1.5), the cell slide was inverted on a glass slide and the cells were observed.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAnnexin V-APC\/DAPI Apoptosis Detection Kit can detect the apoptosis of suspended cells and adherent cells.\u003cbr\u003eAnnexin V is a calcium-dependent phosphatidylserine binding protein with high affinity for phosphatidylserine PS, and Annexin V-APC can bind to the cell membrane of early apoptotic cells through PS exposed extracellularly. Apoptosis can be detected by flow cytometry or fluorescence microscopy.\u003cbr\u003eDue to late apoptosis or loss of membrane integrity in necrotic cells, 4 ', 6-Diamidino-2-phenylindole dihydrochloride (4', 6-Diamidino-2-Phenylindole, DAPI) can specifically bind to double-stranded DNA and produce strong fluorescence. When used in conjunction with AnnexinV, it can distinguish cells in different apoptotic stages.\u003cbr\u003e\u003cstrong\u003eProduct composition:\u003c\/strong\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 77.3056%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 26.3605%; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 15.3795%; text-align: center;\"\u003e20T\u003c\/td\u003e\n\u003ctd style=\"width: 14.0618%; text-align: center;\"\u003e50T\u003c\/td\u003e\n\u003ctd style=\"width: 13.4762%; text-align: center;\"\u003e100T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 26.3605%; text-align: center;\"\u003eAnnexin V-APC\u003c\/td\u003e\n\u003ctd style=\"width: 15.3795%; text-align: center;\"\u003e100 μL\u003c\/td\u003e\n\u003ctd style=\"width: 14.0618%; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 13.4762%; text-align: center;\"\u003e500 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 26.3605%; text-align: center;\"\u003e1X Binding Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 15.3795%; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 14.0618%; text-align: center;\"\u003e25mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.4762%; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 26.3605%; text-align: center;\"\u003eDAPI\u003c\/td\u003e\n\u003ctd style=\"width: 15.3795%; text-align: center;\"\u003e100 μL\u003c\/td\u003e\n\u003ctd style=\"width: 14.0618%; text-align: center;\"\u003e250 μL\u003c\/td\u003e\n\u003ctd style=\"width: 13.4762%; text-align: center;\"\u003e500 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. It should be tested as soon as possible after dyeing. Too long time may lead to increased number of apoptotic or necrotic cells.\u003cbr\u003e2. When detecting adherent cells, the suspension cells produced after apoptosis induction should be collected and detected together with the adherent cells collected subsequently.\u003cbr\u003e3. Try to avoid mechanical damage caused by digestive adherent cells. At the same time, the digestive juice of pancreatic enzymes should be as free of EDTA as possible, as EDTA affects the binding of Annexin V to phosphatidylserine.\u003cbr\u003e4. If pancreatic enzyme containing EDTA is used, the cells should be thoroughly cleaned after collection to ensure that EDTA is removed.\u003cbr\u003e5. Fluorescent substances are easy to quench. When performing fluorescence observation, try to shorten the observation time, and try to pay attention to light preservation during operation and storage.\u003cbr\u003e6. For your safety and health, please wear a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at 2-8 °C, do not freeze, shelf life 1 year.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"20T","offer_id":43214314111051,"sku":"abs50004-20T","price":183.0,"currency_code":"USD","in_stock":true},{"title":"50T","offer_id":41683118358603,"sku":"abs50004-50T","price":285.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41683118391371,"sku":"abs50004-100T","price":439.0,"currency_code":"USD","in_stock":true}]},{"product_id":"tunel-apoptosis-detection-kit","title":"Biotin TUNEL Apoptosis Detection Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e1. Experimental materials (self-prepared)\u003c\/strong\u003e\u003cbr\u003ePBS buffer (1 ×, pH ~ 7.4)\u003cbr\u003e0.2% Triton X-100 (formulated in PBS)\u003cbr\u003eReagents related to paraffin section processing\u003cbr\u003e4% Paraformaldehyde (PBS formulated)\u003cbr\u003eImmunohistochemical pen\u003cbr\u003e0.3% H\u003csub\u003e2\u003c\/sub\u003eO\u003csub\u003e2\u003c\/sub\u003e(Freshly formulated in PBS)\u003cbr\u003eNeutral resin\u003cbr\u003eddH\u003csub\u003e2\u003c\/sub\u003eO\u003cbr\u003e\u003cstrong\u003e2. Experimental design\u003c\/strong\u003e\u003cbr\u003eA. Positive control (optional):\u003cbr\u003ePositive control slides were prepared by DNase I treatment. DNase I can digest single-stranded or double-stranded DNA to produce monodeoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides endonucleases, artificially causing apoptosis.  \u003cbr\u003eB. Negative control (optional):\u003cbr\u003eBiotin TUNEL Reaction Buffer without TdT Enzyme was used with ddH\u003csub\u003e2\u003c\/sub\u003eO replaces TdT Enzyme.  \u003cbr\u003eC. Experimental treatment group.  \u003cbr\u003eD. Experimental control group.  \u003cbr\u003e\u003cstrong\u003e3. Experimental steps\u003c\/strong\u003e\u003cbr\u003e\u003cstrong\u003e1. Sample preparation:\u003c\/strong\u003e \u003cbr\u003e(1) For adherent cells or cell smears\u003cbr\u003ea. PBS wash once.  \u003cbr\u003eNote: If you are worried that the cells of the cell smear will not stick firmly, you can dry the sample to make the cells stick firmly.  \u003cbr\u003eb. Fixation: Add an appropriate amount of 4% paraformaldehyde (prepared with PBS) and fix at room temperature for 30 minutes. PBS wash twice.  \u003cbr\u003ec. Permeability: Add an appropriate amount of 0.2% Triton X-100 (prepared in PBS) and permeate at room temperature for 20 minutes. PBS wash twice.  \u003cbr\u003ed. Blocking: add about 100 μL of 0.3% H per well\u003csub\u003e2\u003c\/sub\u003eO\u003csub\u003e2\u003c\/sub\u003eSolution (freshly prepared with PBS), allowed to fully cover the cells, blocked at room temperature and protected from light for 30 min to inactivate the endogenous catalase in the cells, and then washed twice with PBS.  \u003cbr\u003ee. Go to step 2. TUNEL reaction.  \u003cbr\u003e(2) For suspended cells or cell suspensions\u003cbr\u003eA. Cells were collected (3-5 × 106 cells), centrifuged at 1000 rpm for 5 min, and washed twice with PBS.  \u003cbr\u003eb. Fixation: Add an appropriate amount of 4% paraformaldehyde (prepared with PBS) to fully resuspend the cells, and fix at 4 ℃ for 30 minutes. Centrifuge at 2000 rpm for 5 min and wash twice with PBS.  \u003cbr\u003ec. Permeability: Add an appropriate amount of 0.2% Triton X-100 (prepared in PBS) and permeate at room temperature for 20 minutes. Centrifuge at 2000 rpm for 5 min and wash twice with PBS.  \u003cbr\u003ed. Blocking: add about 100 μL of 0.3% H per well\u003csub\u003e2\u003c\/sub\u003eO\u003csub\u003e2\u003c\/sub\u003eSolution (freshly prepared with PBS), resuspended cells were gently aspirated, blocked at room temperature and protected from light for 30 minutes to inactivate endogenous catalase in cells, and then washed twice with PBS.  \u003cbr\u003ee. Go to step 2. TUNEL reaction.  \u003cbr\u003e(3) Paraffin tissue section\u003cbr\u003eA. Dewaxing and hydration: Section samples are sequentially placed into xylene I (10 min) → xylene II (10 min) → 100% ethanol I (5 min) → 100% ethanol II (5 min) → 95% ethanol (5 min) → 90% ethanol (5 min) → 80% ethanol (5 min) → 70% ethanol (5 min) → ddH\u003csub\u003e2\u003c\/sub\u003eO Rinse for 5 minutes and rinse twice.  \u003cbr\u003eNote: Xylene is toxic and volatile, please do this in a fume hood.  \u003cbr\u003eb. Use filter paper to dry the liquid around the section sample, and circle the sample outline with an immunohistochemical pen to facilitate downstream transparency and marking.  \u003cbr\u003eNote: If the outline circle of immunohistochemical strokes is found to be destroyed in subsequent experimental operations, it is necessary to make up the drawing in time.  \u003cbr\u003ec. Permeability: Dilute 2 mg\/mL Proteinase K solution with PBS to a final concentration of 20 µ g\/mL at a ratio of 1: 100, add 100 µ L dropwise to each sample so that the solution covers the entire sample area and incubate at 20-37 ℃ for 20 min.  \u003cbr\u003eNote: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagent in the subsequent step can fully enter the nucleus for reaction, and improve the labeling efficiency. Too long incubation time will increase the risk of tissue sections falling off the carrier slice in subsequent washing steps, and too short incubation time may cause insufficient permeability treatment and affect labeling efficiency. In order to obtain better results, the concentration, incubation time and temperature of Proteinase K need to be optimized according to different types of tissue samples.  \u003cbr\u003ed. Sections were rinsed twice with PBS for 5 min each time.  \u003cbr\u003eNote: Proteinase K must be cleaned during this step, otherwise it will seriously interfere with the subsequent labeling reaction.  \u003cbr\u003ee. Blocking: add an appropriate amount of 0.3% H\u003csub\u003e2\u003c\/sub\u003eO\u003csub\u003e2\u003c\/sub\u003eThe solution (freshly prepared in PBS) was incubated at room temperature for 30 min to inactivate the endogenous catalase in the sections.  \u003cbr\u003eF. Rinse the sections twice with PBS for 5 min each time, aspirate the excess liquid with filter paper, and place the processed sample in a wet box to keep it moist.  \u003cbr\u003eg. Go to step 2. TUNEL reaction.  \u003cbr\u003e(4) Frozen tissue section\u003cbr\u003eA. Fixation: Take out the frozen sections and warm them to room temperature. Add an appropriate amount of 4% paraformaldehyde (prepared with PBS) and fix it at room temperature for 30 minutes. PBS was rinsed twice for 10 min each time.  \u003cbr\u003eNote: If you are worried that formaldehyde will not be cleaned clean, it will affect the final dyeing effect. After formaldehyde fixation is completed, an appropriate amount of 2 mg\/mL glycine can be added to wash for 10 minutes to neutralize the residual fixative, and then wash with PBS.  \u003cbr\u003eb. Use filter paper to dry the liquid around the section sample, and circle the sample outline with an immunohistochemical pen to facilitate downstream transparency and marking.  \u003cbr\u003eNote: If the outline circle of immunohistochemical strokes is found to be destroyed in subsequent experimental operations, it is necessary to make up the drawing in time.  \u003cbr\u003ec. Permeability: Dilute 2 mg\/mL Proteinase K solution with PBS to a final concentration of 20 µ g\/mL at a ratio of 1: 100, add 100 µ L dropwise to each sample so that the solution covers the entire sample area and incubate at 20-37 ℃ for 20 min.  \u003cbr\u003eNote: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagent in the subsequent step can fully enter the nucleus for reaction, and improve the labeling efficiency. Too long incubation time will increase the risk of tissue sections falling off the carrier slice in subsequent washing steps, and too short incubation time may cause insufficient permeability treatment and affect labeling efficiency. In order to obtain better results, the concentration, incubation time and temperature of Proteinase K need to be optimized according to different types of tissue samples.  \u003cbr\u003ed. Sections were rinsed twice with PBS for 5 min each time.  \u003cbr\u003eNote: Proteinase K must be cleaned during this step, otherwise it will seriously interfere with the subsequent labeling reaction.  \u003cbr\u003ee. Blocking: add an appropriate amount of 0.3% H\u003csub\u003e2\u003c\/sub\u003eO\u003csub\u003e2\u003c\/sub\u003eThe solution (freshly prepared in PBS) was incubated at room temperature for 30 min to inactivate the endogenous catalase in the sections.  \u003cbr\u003eF. Rinse the sections twice with PBS for 5 min each time, suck off the excess liquid with filter paper, and place the treated sections in a wet box to keep them moist.  \u003cbr\u003eg. Go to step 2. TUNEL reaction.  \u003cbr\u003e(5) Positive treatment (only positive control is subjected to this step, other samples are directly subjected to TUNEL reaction step)\u003cbr\u003eA. Using ddH at a ratio of 1: 10\u003csub\u003e2\u003c\/sub\u003e0 10 × DNase I Buffer was diluted to 1 × DNase I Buffer for later use.\u003cbr\u003eb. Add 100 µ L of 1 × DNase I Buffer dropwise to the processed sample covering the entire sample area and equilibrate at room temperature for 5 min.  \u003cbr\u003ec. DNase I (2 U\/μL) was diluted 1: 100 with 1 × DNase I Buffer to a final concentration of 20 U\/mL of the working solution.  \u003cbr\u003ed. Discard the Buffer, add 100 μL of DNase I working solution at a concentration of 20 U\/mL, and incubate at room temperature for 10 min.  \u003cbr\u003ee. Discard the DNase I working solution and wash twice with PBS.  \u003cbr\u003ef. Go to step 2. TUNEL reaction.  \u003cbr\u003e\u003cstrong\u003e2. TUNEL reaction\u003c\/strong\u003e\u003cbr\u003e(1) Prepare TUNEL reaction solution (ready for use):\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 75.0329%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 44.707%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 16.3356%; text-align: center;\"\u003e1 sample\u003c\/td\u003e\n\u003ctd style=\"width: 15.6352%; text-align: center;\"\u003e5 samples\u003c\/td\u003e\n\u003ctd style=\"width: 15.9854%; text-align: center;\"\u003e10 samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 44.707%; text-align: center;\"\u003eTdT enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 16.3356%; text-align: center;\"\u003e1 μL\u003c\/td\u003e\n\u003ctd style=\"width: 15.6352%; text-align: center;\"\u003e5 μL\u003c\/td\u003e\n\u003ctd style=\"width: 15.9854%; text-align: center;\"\u003e10 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 44.707%; text-align: center;\"\u003eBiotin TUNEL Reaction Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 16.3356%; text-align: center;\"\u003e49 μL\u003c\/td\u003e\n\u003ctd style=\"width: 15.6352%; text-align: center;\"\u003e2145 μL\u003c\/td\u003e\n\u003ctd style=\"width: 15.9854%; text-align: center;\"\u003e490 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 44.707%; text-align: center;\"\u003eTotal TUNEL reaction solution volume\u003c\/td\u003e\n\u003ctd style=\"width: 16.3356%; text-align: center;\"\u003e50 μL\u003c\/td\u003e\n\u003ctd style=\"width: 15.6352%; text-align: center;\"\u003e250 μL\u003c\/td\u003e\u0026lt; td style = \"width: 15.9854%; text-align: center; \"\u0026gt; 500 μL\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e(2) Add 50μL of TUNEL reaction solution to each sample so that the reaction solution evenly covers the sample. Incubate at 37 °C for 60 min.  \u003cbr\u003eNote: 50μL TUNEL reaction solution is suitable for smear, section or 96-well plate (other different well plates can appropriately adjust the volume of TUNEL reaction solution to cover cells).  \u003cbr\u003eIf the sample to be tested is a smear, section or in a 24-well plate, 12-well plate or 6-well plate, you can use an anti-evaporation film, or try to use a ziplock bag or other appropriate material to cut it into a round plastic sheet slightly smaller than the well. Add TUNEL reaction solution dropwise and cover it on the sample, which can prevent the evaporation of TUNEL reaction solution and make the TUNEL reaction solution evenly cover the sample.  \u003cbr\u003e(3) Discard the TUNEL reaction solution and wash it twice with PBS.  \u003cbr\u003e\u003cstrong\u003e3. Preparation of Streptavidin-HRP working solution and DAB color development solution\u003c\/strong\u003e\u003cbr\u003e(1) Preparation of Streptavidin-HRP working solution (ready to use):\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 69.7766%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.8099%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 17.1394%; text-align: center;\"\u003e1 sample\u003c\/td\u003e\n\u003ctd style=\"width: 17.7044%; text-align: center;\"\u003e5 samples\u003c\/td\u003e\n\u003ctd style=\"width: 16.0095%; text-align: center;\"\u003e10 samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.8099%; text-align: center;\"\u003e\u003cp\u003eStreptavidin-HRP\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 17.1394%; text-align: center;\"\u003e1 ul\u003c\/td\u003e\n\u003ctd style=\"width: 17.7044%; text-align: center;\"\u003e5 ul\u003c\/td\u003e\n\u003ctd style=\"width: 16.0095%; text-align: center;\"\u003e10 ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.8099%; text-align: center;\"\u003e\u003cp\u003eStreptavidin-HRP dilution\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 17.1394%; text-align: center;\"\u003e49 ul\u003c\/td\u003e\n\u003ctd style=\"width: 17.7044%; text-align: center;\"\u003e245 ul\u003c\/td\u003e\n\u003ctd style=\"width: 16.0095%; text-align: center;\"\u003e490 ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 41.8099%; text-align: center;\"\u003e\u003cp\u003eTotal Volume of Streptavidin-HRP Working Fluid\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 17.1394%; text-align: center;\"\u003e50 ul\u003c\/td\u003e\n\u003ctd style=\"width: 17.7044%; text-align: center;\"\u003e250 ul\u003c\/td\u003e\n\u003ctd style=\"width: 16.0095%; text-align: center;\"\u003e500 ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e(2) Preparation of DAB chromogenic solution (ready for use):\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 69.251%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 43.4695%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 17.663%; text-align: center;\"\u003e1 sample\u003c\/td\u003e\n\u003ctd style=\"width: 16.1448%; text-align: center;\"\u003e5 samples\u003c\/td\u003e\n\u003ctd style=\"width: 15.3859%; text-align: center;\"\u003e10 samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 43.4695%; text-align: center;\"\u003eDAB chromogenic solution A\u003c\/td\u003e\n\u003ctd style=\"width: 17.663%; text-align: center;\"\u003e5 ul\u003c\/td\u003e\n\u003ctd style=\"width: 16.1448%; text-align: center;\"\u003e25 ul\u003c\/td\u003e\n\u003ctd style=\"width: 15.3859%; text-align: center;\"\u003e50 ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 43.4695%; text-align: center;\"\u003eDAB chromogenic solution B\u003c\/td\u003e\n\u003ctd style=\"width: 17.663%; text-align: center;\"\u003e42.5 ul\u003c\/td\u003e\n\u003ctd style=\"width: 16.1448%; text-align: center;\"\u003e212.5 ul\u003c\/td\u003e\n\u003ctd style=\"width: 15.3859%; text-align: center;\"\u003e425 ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 43.4695%; text-align: center;\"\u003eDAB chromogenic solution C\u003c\/td\u003e\n\u003ctd style=\"width: 17.663%; text-align: center;\"\u003e2.5 ul\u003c\/td\u003e\n\u003ctd style=\"width: 16.1448%; text-align: center;\"\u003e12.5 ul\u003c\/td\u003e\n\u003ctd style=\"width: 15.3859%; text-align: center;\"\u003e25 ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 43.4695%; text-align: center;\"\u003eTotal volume of DAB chromogenic solution\u003c\/td\u003e\n\u003ctd style=\"width: 17.663%; text-align: center;\"\u003e50 ul\u003c\/td\u003e\n\u003ctd style=\"width: 16.1448%; text-align: center;\"\u003e250 ul\u003c\/td\u003e\n\u003ctd style=\"width: 15.3859%; text-align: center;\"\u003e500ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cstrong\u003e4. Sample color development\u003cbr\u003e\u003c\/strong\u003e(1) Add 50 μL of Streptavidin-HRP working solution to each sample and incubate at 37 ℃ for 30 min.\u003cbr\u003eNote: 50μL Streptavidin-HRP working solution is suitable for smear, section or 96-well plate (other different well plates can appropriately adjust the volume of Streptavidin-HRP working solution to cover cells). To prevent evaporation of the Streptavidin-HRP working fluid, it is recommended to coat the samples with an anti-evaporation film.\u003cbr\u003e(2) Discard the Streptavidin-HRP working solution and wash it twice with PBS.\u003cbr\u003e(3) Add 50μL of DAB color development solution to each sample, incubate at room temperature for 5 minutes or control the staining time under a microscope according to the color development.\u003cbr\u003eNote: If the color development is very strong, the color development can be stopped in less than 5 minutes. If the color development is very weak, the color development time can be appropriately extended, or even the color development can be performed overnight.\u003cbr\u003e(4) Discard the DAB color solution and wash it twice with PBS.\u003cbr\u003e(5) (Optional) Add an appropriate amount of hematoxylin staining solution or methyl green staining solution for nucleus staining. The staining solution was discarded and washed twice with PBS.\u003cbr\u003e(6) (Optional) Slice sealing: Immerse the slices in pure water, 70% ethanol, 80% ethanol, 90% ethanol, 95% ethanol and absolute ethanol for 5 minutes in turn, and finally put the slice samples in a dyeing tank. Soak in fresh xylene, and transparent treatment twice for 5 minutes each time. After the dehydration was complete, the liquid around the sections was wiped away, 50 μL of neutral resin was added dropwise to each sample, a coverslip was covered, and the coverslip was gently tapped with the blunt end of the tweezers to remove air bubbles to complete the sealing.\u003cbr\u003e(7) Use filter paper to suck off excess liquid, add 100μL PBS to the sample area to keep the sample moist, and immediately analyze the sample under an optical microscope.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eExperiment Reagent Instrument\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003ePBS,Paraformaldehyde,Triton X-100,Methanol,H2O2,Hematoxylin (optional)\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eWhen cells undergo apoptosis, some DNA endonucleases will be activated. These endonucleases will cut off the genomic DNA between nucleosomes and produce DNA fragments of 180 bp to 200 bp, which shows the specific Ladder Ladder map presented in agarose gel electrophoresis. When genomic DNA double-stranded or single-stranded breaks, a large number of sticky 3 '-OH terminals will occur, which can be combined with Biotin-dUTP under the catalysis of deoxyribonucleotide terminal transferase (TdT), so as to directly detect apoptotic cells through light microscopy. This method is called Terminal-deoxynucleotidyl transfer mediated nick end labeling (TUNEL). Since normal or proliferating cells have little DNA breakage, there is no 3 '-OH formation and little can be stained. Tunel method can stain complete single apoptotic nuclei or apoptotic bodies in situ, accurately reflect the typical biochemical and morphological characteristics of apoptosis, and can detect a very small number of apoptotic cells. Therefore, it is widely used in the research of apoptosis.  \u003cbr\u003eThe kit has a wide range of applications and can be used to detect the apoptosis of cells in frozen or paraffin sections, and can also detect the apoptosis of cultured adherent cells or suspended cells. It can selectively detect apoptotic cells, but not necrotic cells or cells with DNA strand breaks caused by irradiation and drug treatment.  \u003cbr\u003e\u003cstrong\u003eComponent Information:\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 73.3237%; height: 220px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eSerial number\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eComponent Name\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e50T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eBiotin TUNEL Reaction Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e2 × 1. 25 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eTdT Enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e50uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eStreptavidin-HRP\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e50uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eD\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eStreptavidin-HRP dilution\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e2 × 1. 25 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eE\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eDAB chromogenic solution A\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e250uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eF\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eDAB chromogenic solution B\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e2 × 1. 25 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eG\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eDAB chromogenic solution C\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e125uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eH\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eProteinase K (2 mg\/mL)\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e100uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eI\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; height: 22px; text-align: center;\"\u003eDNase I (2 U\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; height: 22px; text-align: center;\"\u003e13uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 8.30756%; text-align: center;\"\u003eJ\u003c\/td\u003e\n\u003ctd style=\"width: 35.956%; text-align: center;\"\u003e10 × DNase I Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 10.2887%; text-align: center;\"\u003e260uL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. For your safety and health, please wear a lab coat and disposable gloves.\u003cbr\u003e2. Sodium azide has an inhibitory effect on HRP. Do not use reagents containing sodium azide in the experiment.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThis product should be stored at-20 ℃ protected from light; Avoid repeated freezing and thawing. This product can be stored for up to 12 months under recommended conditions.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50T","offer_id":41683111215179,"sku":"abs50022-50T","price":508.0,"currency_code":"USD","in_stock":true}]},{"product_id":"ao-staining-solution","title":"AO staining solution (1 mg\/ml)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Collect cells (fix the cells first when using flow cytometry), wash the cells once with PBS, count the cells, and adjust the cell concentration to 10\u003csup\u003e6\u003c\/sup\u003e\/ml. 2. Take an appropriate amount of cell suspension and add Acridine Orange Stain (1 mg\/ml) to make the final AO concentration between 8.5 and 17 μg\/ml. Mix gently. \u003cbr\u003e3. Stain at room temperature in the dark for 15 minutes. Place the cells onto a glass slide and cover slip with a glass cover slip or analyze on a flow cytometer. \u003cbr\u003e4. Observe under a fluorescence microscope (excitation filter wavelength 488nm, blocking filter wavelength 515nm), count, and photograph. \u003cbr\u003e\u003cstrong\u003eStaining Results:\u003c\/strong\u003e\u003cbr\u003eNormal cells: Cells are uniformly stained with a yellow-green fluorescence. \u003cbr\u003eApoptotic cells: Chromatin is condensed, and the nucleus is fragmented into punctate shapes, which are stained as dense, intensely stained green granules of varying sizes.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eAppearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eSolution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAcridine Orange is a tricyclic heteroaromatic dye that can label DNA and RNA, and is a metachromatic fluorescent dye. This dye is membrane-permeable and can penetrate cell membranes to stain nuclear DNA and RNA. Therefore, AO is often used for detecting intracellular DNA and RNA. AO binds to nucleic acids in two main ways: 1. Intercalation, where AO intercalates between base pairs in double-stranded nucleic acids. This binding mode primarily occurs with DNA, with a fluorescence emission peak at 530 nm and green fluorescence upon excitation. 2. Electrostatic attraction, where the positively charged AO binds to the negatively charged phosphate groups of single-stranded nucleic acids through electrostatic attraction. This binding mode primarily occurs with RNA, with a fluorescence emission peak at 640 nm and red fluorescence upon excitation. Small amounts of binding can result in orange or red fluorescence. Therefore, acridine orange appears green when embedded in double-stranded DNA, and emits orange or red fluorescence when bound to single-stranded DNA or RNA. \u003cbr\u003e        AO staining solution should be diluted to an appropriate concentration before use. After staining, observation under a fluorescence microscope reveals that AO can penetrate normal cell membranes, causing the cell nucleus to exhibit a uniform green or yellow-green fluorescence. In apoptotic cells, chromatin condenses or breaks into fragments of varying sizes, forming apoptotic bodies. AO stains these cells with dense, intense yellow-green fluorescence or yellow-green fragments, while the yellow fluorescence of necrotic cells weakens or even disappears. AO staining is often combined with EB staining for double staining, as EB only stains dead cells, causing them to produce orange-yellow fluorescence. This allows for the distinction between normal, apoptotic, and necrotic cells.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. This product does not contain a membrane permeabilizing agent and is rarely used alone. \u003cbr\u003e 2. AO staining is often used in combination with EB staining to distinguish normal cells from apoptotic cells and necrotic cells. \u003cbr\u003e 3. Centrifugation in a low-temperature centrifuge is more effective. \u003cbr\u003e 4. During operation, be careful to minimize exposure of the reagent to strong light. \u003cbr\u003e 5. For your safety and health, please wear a lab coat and disposable gloves during operation.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eConcentration\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1mg\/ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e2-8℃, protect from light, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"10mL","offer_id":41683112853579,"sku":"abs9735-10mL","price":19.0,"currency_code":"USD","in_stock":true}]},{"product_id":"tunel-fitc-abs50058","title":"TUNEL apoptosis kit (red fluorescence)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e1. Experimental materials (self-prepared)\u003cbr\u003e\u003c\/strong\u003ePBS buffer (1 ×, pH ~ 7.4)\u003cbr\u003e0.2% Triton X-100 (formulated in PBS)\u003cbr\u003e0.1% Triton X-100 in PBS containing 5 mg\/mL BSA\u003cbr\u003e4% Paraformaldehyde (PBS formulated)\u003cbr\u003eImmunohistochemical pen\u003cbr\u003eDewaxing Solvent (Paraffin Section Sample)\u003cbr\u003eReagents related to paraffin section processing\u003cbr\u003eAnti-fluorescence quenching sealing tablet\u003cbr\u003eddH\u003csub\u003e2\u003c\/sub\u003eO\u003cbr\u003e\u003cstrong\u003e2. Experimental design\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eA. Positive control:\u003cbr\u003e\u003c\/strong\u003ePositive control slides were prepared by DNase I treatment. DNase I can digest single-stranded or double-stranded DNA to produce monodeoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides endonucleases, artificially causing apoptosis.\u003cbr\u003e\u003cstrong\u003eB. Negative control:\u003cbr\u003e\u003c\/strong\u003eUsing TUNEL Reaction Buffer without TdT Enzyme, using ddH\u003csub\u003e2\u003c\/sub\u003eO replaces TdT Enzyme.\u003cbr\u003e\u003cstrong\u003eC. Experimental treatment group.\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eD. Experimental control group.\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e3. Experimental steps:\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e1. Sample preparation:\u003cbr\u003e\u003c\/strong\u003e(1) For adherent cells or cell smears\u003cbr\u003ea. PBS wash once.\u003cbr\u003eNote: If you are worried that the cells of the cell smear will not stick firmly, you can dry the sample to make the cells stick firmly.\u003cbr\u003eb. Fixation: Add an appropriate amount of 4% paraformaldehyde (formulated in PBS) and fix at 4 °C for 30 min. PBS wash twice.\u003cbr\u003ec. Permeability: Add an appropriate amount of 0.2% Triton X-100 (prepared in PBS) and permeate at room temperature for 20 minutes. PBS wash twice.\u003cbr\u003ed. Transfer steps\u003cstrong\u003e2. TUNEL\u003c\/strong\u003e\u003cstrong\u003ereaction\u003c\/strong\u003e。\u003c\/p\u003e\n\u003cp\u003e(2) For suspended cells or cell suspensions\u003cbr\u003ea. Cells were harvested (3-5 × 10\u003csup\u003e6\u003c\/sup\u003eCells), centrifuged at 1000 rpm for 5 min, and washed twice with PBS.\u003cbr\u003eb. Fixation: Add an appropriate amount of 4% paraformaldehyde (prepared with PBS) to fully resuspend the cells, and fix at 4 ℃ for 30 minutes. Centrifuge at 2000 rpm for 5 min and wash twice with PBS.\u003cbr\u003ec. Permeability: Add an appropriate amount of 0.2% Triton X-100 (prepared in PBS) and permeate at room temperature for 20 minutes. Centrifuge at 2000 rpm for 5 min and wash twice with PBS.\u003cbr\u003ed. Transfer steps\u003cstrong\u003e2. TUNEL\u003c\/strong\u003e\u003cstrong\u003ereaction\u003c\/strong\u003e。\u003c\/p\u003e\n\u003cp\u003e(3) Paraffin tissue section\u003cbr\u003eA. Dewaxing and hydration: Section samples are sequentially placed into xylene I (10 min) → xylene II (10 min) → 100% ethanol I (5 min) → 100% ethanol II (5 min) → 95% ethanol (5 min) → 90% ethanol (5 min) → 80% ethanol (5 min) → 70% ethanol (5 min) → ddH\u003csub\u003e2\u003c\/sub\u003eO Rinse for 5 minutes and rinse twice.\u003cbr\u003eNote: Xylene is toxic and volatile, please do this in a fume hood.\u003cbr\u003eb. Use filter paper to dry the liquid around the section sample, and circle the sample outline with an immunohistochemical pen to facilitate downstream transparency and marking.\u003cbr\u003eNote: If the outline circle of immunohistochemical strokes is found to be destroyed in subsequent experimental operations, it is necessary to make up the drawing in time.\u003cbr\u003ec. Permeability: Dilute 2 mg\/mL Proteinase K solution with PBS to a final concentration of 20 µ g\/mL at a ratio of 1: 100, add 100 µ L dropwise to each sample so that the solution covers the entire sample area and incubate at 20-37 ℃ for 20 min.\u003cbr\u003eNote: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagent in the subsequent step can fully enter the nucleus for reaction, and improve the labeling efficiency. Too long incubation time will increase the risk of tissue sections falling off the carrier slice in subsequent washing steps, and too short incubation time may cause insufficient permeability treatment and affect labeling efficiency. In order to obtain better results, the concentration, incubation time and temperature of Proteinase K need to be optimized according to different types of tissue samples.\u003cbr\u003ed. Sections were rinsed twice with PBS for 5 min each time, excess liquid was aspirated with filter paper, and processed samples were kept moist in a wet box.\u003cbr\u003eNote: Proteinase K must be cleaned during this step, otherwise it will seriously interfere with the subsequent labeling reaction.\u003cbr\u003ee. Transfer steps\u003cstrong\u003e2. TUNEL\u003c\/strong\u003e\u003cstrong\u003ereaction\u003c\/strong\u003e。\u003c\/p\u003e\n\u003cp\u003e(4) Frozen tissue section\u003cbr\u003ea. Fixation: Remove the frozen sections and warm them to room temperature. Add an appropriate amount of 4% paraformaldehyde (prepared with PBS) and fix it at room temperature for 30 minutes. PBS was rinsed twice for 10 min each time.\u003cbr\u003eNote: If you are worried that formaldehyde will not be cleaned clean, it will affect the final dyeing effect. After formaldehyde fixation is completed, an appropriate amount of 2 mg\/mL glycine can be added to wash for 10 minutes to neutralize the residual fixative, and then wash with PBS.\u003cbr\u003eb. Use filter paper to dry the liquid around the section sample, and circle the sample outline with an immunohistochemical pen to facilitate downstream transparency and marking.\u003cbr\u003eNote: If the outline circle of immunohistochemical strokes is found to be destroyed in subsequent experimental operations, it is necessary to make up the drawing in time.\u003cbr\u003ec. Permeability: Dilute 2 mg\/mL Proteinase K solution with PBS to a final concentration of 20 µ g\/mL at a ratio of 1: 100, add 100 µ L dropwise to each sample so that the solution covers the entire sample area and incubate at 20-37 ℃ for 20 min.\u003cbr\u003eNote: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagent in the subsequent step can fully enter the nucleus for reaction, and improve the labeling efficiency. Too long incubation time will increase the risk of tissue sections falling off the carrier slice in subsequent washing steps, and too short incubation time may cause insufficient permeability treatment and affect labeling efficiency. In order to obtain better results, the concentration, incubation time and temperature of Proteinase K need to be optimized according to different types of tissue samples.\u003cbr\u003ed. Sections were rinsed twice with PBS for 5 min each time, excess liquid was aspirated with filter paper, and processed samples were kept moist in a wet box.\u003cbr\u003eNote: Proteinase K must be cleaned during this step, otherwise it will seriously interfere with the subsequent labeling reaction.\u003cbr\u003ee. Transfer steps\u003cstrong\u003e2. TUNEL\u003c\/strong\u003e\u003cstrong\u003ereaction\u003c\/strong\u003e。\u003c\/p\u003e\n\u003cp\u003e(5) Positive treatment (only positive control is subjected to this step, other samples are directly subjected to TUNEL reaction step)\u003cbr\u003ea. With ddH at a ratio of 1: 10\u003csub\u003e2\u003c\/sub\u003e0 10 × DNase I Buffer was diluted to 1 × DNase I Buffer for later use.\u003cbr\u003eb. Add 100 µ L of 1 × DNase I Buffer dropwise to the processed sample covering the entire sample area and equilibrate at room temperature for 5 min.\u003cbr\u003ec. DNase I (2 U\/μL) was diluted 1: 100 with 1 × DNase I Buffer to a final concentration of 20 U\/mL of the working solution.\u003cbr\u003ed. Discard the Buffer, add 100 μL of DNase I working solution at a concentration of 20 U\/mL, and incubate at room temperature for 10 min.\u003cbr\u003ee. Discard the DNase I working solution and wash twice with PBS.\u003cbr\u003ef. Transfer steps\u003cstrong\u003e2. TUNEL\u003c\/strong\u003e\u003cstrong\u003ereaction\u003c\/strong\u003e。\u003cbr\u003e\u003cstrong\u003e2. TUNEL\u003c\/strong\u003e\u003cstrong\u003ereaction\u003cbr\u003e\u003c\/strong\u003e(1) Prepare TUNEL reaction solution (ready for use):\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 83.9685%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e1 sample\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e5 samples\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e10 samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003eTdT Enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003cp\u003e1 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\u003cp\u003e5 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\u003cp\u003e10 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e\u003cp\u003eTUNEL Reaction Buffer\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003cp\u003e49 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\u003cp\u003e245 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\u003cp\u003e490 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e\u003cp\u003eTotal TUNEL reaction solution volume\u003c\/p\u003e\u003c\/td\u003e\u0026lt; td style = \"width: 14.7558%; text-align: center; \"\u0026gt;\u003cp\u003e50 μL\u003c\/p\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\u003cp\u003e250 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\u003cp\u003e500 μL\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e\u003cp\u003e(2) For adherent cells, cell smears or tissue sections\u003cbr\u003eA. Add 50 μL of TUNEL reaction solution to each sample so that the reaction solution evenly covers the sample. Incubate at 37 ℃ for an appropriate time in the dark (the recommended staining time for cells is 30min-1h, and the recommended staining time for tissues is 2h).\u003cbr\u003eNote: 50μL TUNEL reaction solution is suitable for smear, section or 96-well plate (other different well plates can appropriately adjust the volume of TUNEL reaction solution to cover cells).\u003cbr\u003eIf the sample to be tested is a smear, section or in a 24-well plate, 12-well plate or 6-well plate, you can use an anti-evaporation film, or try to use a ziplock bag or other appropriate material to cut it into a round plastic sheet slightly smaller than the well. Add TUNEL reaction solution dropwise and cover the sample, which can prevent the evaporation of TUNEL reaction solution and make the TUNEL reaction solution evenly cover the sample.\u003cbr\u003eb. Discard the TUNEL reaction solution, rinse it twice with PBS, and then rinse it three times with 0.1% Triton X-100 (formulated in PBS containing 5 mg\/mL BSA) for 5 minutes each time, so that free unreacted markers can be cleaned.\u003cbr\u003ec. (Optional) Add an appropriate amount of DAPI dye solution with a concentration of 5μg\/mL to each sample, and incubate at room temperature and protect from light for 5 minutes. After completion of staining, the DAPI stain solution was discarded and rinsed twice with PBS for 5 min each time.\u003cbr\u003ed. (Optional) Section sealing: Add 50μL of anti-fluorescence quenching sealing tablet dropwise to each sample (anti-fluorescence quenching sealing tablet may not be suitable for some dyes, and it is recommended to conduct a pre-experiment to test the matching before the experiment), cover the coverslip, and gently tap the coverslip with the blunt end of tweezers to remove air bubbles to complete the sealing.\u003cbr\u003ee. Use filter paper to suck off excess liquid, add 100μL PBS to the sample area to keep the sample moist, and immediately observe under a fluorescence microscope.\u003c\/p\u003e\n\u003cp\u003e(3) For suspended cells or cell suspensions\u003cbr\u003ea. Add 50 μL of TUNEL reaction solution to each sample tube to gently resuspend the cells, and incubate at 37 °C in the dark for 30-60 minutes. The cells were gently resuspended with a micropipette every 15 min.\u003cbr\u003eb. Centrifuge at 2000 rpm for 5 minutes, discard the TUNEL reaction solution, add an appropriate amount of 0.1% Triton X-100 (prepared in PBS, containing 5 mg\/mL BSA) to gently resuspend the cells, and wash twice for 5 minutes each time, so that the free unreacted markers can be cleaned.\u003cbr\u003ec. Each sample tube was added with 100 μL of DAPI dye solution with a concentration of 5 μg\/mL, and incubated at room temperature and protected from light for 5 minutes.\u003cbr\u003ed. 400 μL of PBS was added to resuspend cells and immediately detected by flow cytometry or smeared and observed under a fluorescence microscope.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTUNEL Apoptosis Detection Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDetection Type\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eSample types: paraffin section, frozen section, adherent cells and suspension cells.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eWhen cells undergo apoptosis, some DNA endonucleases will be activated. These endonucleases will cut off the genomic DNA between nucleosomes and produce DNA fragments of 180 bp to 200 bp, which shows the specific Ladder Ladder map presented in agarose gel electrophoresis. When genomic DNA double-stranded or single-stranded breaks, a large number of sticky 3 '-OH ends will be produced, which can be combined with 594\/Cy-dUTP under the catalysis of deoxyribonucleotide terminal transferase (TdT), so as to pass through fluorescence microscope or flow cytometry directly detects apoptotic cells. This method is called Terminal-deoxynucleotidyl transfer mediated nick end labeling (TUNEL). Since normal or proliferating cells have little DNA breakage, there is no 3 '-OH formation and little can be stained. Tunel method can stain complete single apoptotic nuclei or apoptotic bodies in situ, accurately reflect the typical biochemical and morphological characteristics of apoptosis, and can detect a very small number of apoptotic cells. Therefore, it is widely used in the research of apoptosis.  \u003cbr\u003eThe kit has a wide range of applications and can be used to detect the apoptosis of cells in frozen or paraffin sections, and can also detect the apoptosis of cultured adherent cells or suspended cells. It can selectively detect apoptotic cells, but not necrotic cells or cells with DNA strand breaks caused by irradiation and drug treatment. The kit takes short time to detect cell apoptosis, only needs one step of staining reaction, and can be detected after washing.\u003cbr\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 77.7072%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eserial number\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e20T\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e50T\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eTUNEL Reaction Buffer (594)\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e1 mL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e2 × 1. 25 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eTdT Enzyme\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e20 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e50 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eProteinase K (2 mg\/mL)\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e40 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e100 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eD\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003eDNase I (2 U\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e5 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e13 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 1.82482%; text-align: center;\"\u003eE\u003c\/td\u003e\n\u003ctd style=\"width: 9.57316%; text-align: center;\"\u003e10 × DNase I Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 4.15853%; text-align: center;\"\u003e100 μL\u003c\/td\u003e\n\u003ctd style=\"width: 3.01781%; text-align: center;\"\u003e260 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Before use, please centrifuge the product to the bottom of the tube instantly, and then conduct subsequent experiments.  \u003cbr\u003e2. When the staining background is heavy or the non-specific staining is obvious, the staining time can be appropriately reduced.  \u003cbr\u003e3. It is recommended to add negative control and positive control group during the experiment.  \u003cbr\u003e4. Please wear a mask and gloves when using component A. If it comes into contact with the skin, please rinse with plenty of water immediately.  \u003cbr\u003e5. Fluorescent dyes all have quenching problems. Please try to avoid light to slow down fluorescence quenching.  \u003cbr\u003e6. For your safety and health, please wear a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eInstructions\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eI. Experimental materials (self supplied) \u003cbr\u003e\u003c\/strong\u003e  PBS buffer (1× , pH~7.4) \u003cbr\u003e0.2% Triton X-100 (prepared in PBS) \u003cbr\u003e0.1% Triton X-100 (prepared in PBS, Containing 5 mg\/mL BSA) \u003cbr\u003e4% paraformaldehyde (PBS) \u003cbr\u003eimmunohistochemical pen \u003cbr\u003edeparaffinizing solvent (paraffin section samples) \u003cbr\u003ereagents related to paraffin section processing \u003cbr\u003eanti-fluorescence quenching tablet \u003cbr\u003eddH\u003csub\u003e2\u003c\/sub\u003eO\u003cbr\u003e\u003cstrong\u003eII. Experimental design \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eA. Positive control: \u003cbr\u003e\u003c\/strong\u003eDNase I treatment was used to prepare positive control slides. DNase I can digest single - or double-stranded DNA to produce monodeoxynucleotides or single - or double-stranded oligodeoxynucleotides, which artificially causes apoptosis. \u003cbr\u003e\u003cstrong\u003eB. Negative control: \u003cbr\u003e\u003c\/strong\u003eTUNEL Reaction Buffer without TdT Enzyme was used and ddH\u003csub\u003e2\u003c\/sub\u003eO was used instead of TdT Enzyme. \u003cbr\u003e\u003cstrong\u003eC. Experimental treatment groups. \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eD. Experimental control group. \u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003eIII. Experimental procedures :\u003cbr\u003e\u003c\/strong\u003e\u003cstrong\u003e1. Sample preparation :\u003cbr\u003e\u003c\/strong\u003e(1) for adherent cells or cell smears \u003cbr\u003e      a. Rinse 1 time with PBS. \u003cbr\u003e      Note: If there is concern that the cells of the cell smear are not attached firmly, the sample can be dried to make the cells attach more firmly. \u003cbr\u003e      b. Fixed: add appropriate amount of 4% paraformaldehyde (PBS preparation), 4° C fixed for 30 min. Wash twice with PBS. \u003cbr\u003e      c. Permeabilization: Add appropriate amount of 0.2% Triton X-100 (prepared in PBS) and permeabilize for 20 min at room temperature. Rinse twice with PBS. \u003cbr\u003e      d. Turn step \u003cstrong\u003e2. TUNEL \u003c\/strong\u003e\u003cstrong\u003ereaction \u003c\/strong\u003e. \u003c\/p\u003e\n\u003cp\u003e(2) for suspension cells or cell suspensions \u003cbr\u003e      a. Collect cells (3-5× 10\u003csup\u003e6\u003c\/sup\u003ecells), centrifuged at 1000 rpm for 5 min, and washed twice with PBS. \u003cbr\u003e      b. Fixation: Cells were fully resuspended by adding an appropriate amount of 4% paraformaldehyde (prepared in PBS) and fixed at 4 ° C for 30 min. The cells were centrifuged at 2000 rpm for 5 min and washed twice with PBS. \u003cbr\u003e      c. Permeabilization: Add appropriate amount of 0.2% Triton X-100 (prepared in PBS) and permeabilize for 20 min at room temperature. The mixture was centrifuged at 2000 rpm for 5 min and washed twice with PBS. \u003cbr\u003e      d. Turn step \u003cstrong\u003e2. TUNEL \u003c\/strong\u003e\u003cstrong\u003ereaction \u003c\/strong\u003e. \u003c\/p\u003e\n\u003cp\u003e(3) Paraffin tissue sections \u003cbr\u003e      a.  Deparaffinization and hydration: Put the sliced samples in xylene I (10 min) in turn. Xylene II (10 min) → 100% ethanol I (5 min) → 100% ethanol II (5 min) → 95% ethanol (5 min) → 90% ethanol (5 min) → 80% ethanol (5 min) → 70% ethanol (5 min) → ddH\u003csub\u003e2\u003c\/sub\u003eO was rinsed for 5 min and twice. \u003cbr\u003e      Note: Xylene is toxic and volatile, do this in a fume hood. \u003cbr\u003e      b. Blot the liquid around the sliced sample with filter paper, and circle the sample outline with an IHC pen for downstream permeability and labeling. \u003cbr\u003e      Note: If the outline circle of immunohistochemical strokes is found to be destroyed in the subsequent experimental operation, it should be repaired in time. \u003cbr\u003e      c. Permeabilization: at a ratio of 1:100, 2 mg\/mL of Proteinase K solution was diluted with PBS to a final concentration of 20 µ g\/mL, add 100 \u0026amp;micro droppers to each sample; L, so that the solution covered the entire sample area and incubated for 20 min at 20-37 ° C. \u003cbr\u003e      Note: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagents in the subsequent steps can fully enter the nucleus for reaction and improve the labeling efficiency. Too long of the incubation time will increase the risk of the tissue section falling off the carrier plate in the subsequent washing step, and too short of the incubation time may cause inadequate permeability treatment and affect the labeling efficiency. In order to obtain better results, the concentration, incubation time and temperature of Proteinase K should be optimized according to different types of tissue samples. \u003cbr\u003e      d. Rinse the sections twice with PBS for 5 min each time, blot off excess liquid with filter paper, and keep the processed sample moist in a wet box. \u003cbr\u003e      Note: This step must wash the Proteinase K clean, otherwise it will seriously interfere with the subsequent labeling reaction. \u003cbr\u003ee. Turn step \u003cstrong\u003e2. TUNEL \u003c\/strong\u003e\u003cstrong\u003ereaction \u003c\/strong\u003e. \u003c\/p\u003e\n\u003cp\u003e(4) Frozen tissue sections \u003cbr\u003e      a. Fix: Remove frozen sections and return to room temperature. Add appropriate amount of 4% paraformaldehyde (prepared in PBS) and fix at room temperature for 30 min. Rinse twice with PBS for 10 min each time. \u003cbr\u003e      Note: If you worry about formaldehyde cleaning is not clean, affect the final dyeing effect. After formaldehyde fixation is completed, an appropriate amount of 2 mg\/mL glycine can be added to clean for 10 min to neutralize the residual fixative, and then clean with PBS. \u003cbr\u003e      b. Blot the liquid around the sliced sample with filter paper, and circle the sample outline with an IHC pen for downstream permeability and labeling. \u003cbr\u003e      Note: If the outline circle of immunohistochemical strokes is found to be destroyed in the subsequent experimental operation, it should be repaired in time. \u003cbr\u003e      c. Permeabilization: at a ratio of 1:100, 2 mg\/mL of Proteinase K solution was diluted with PBS to a final concentration of 20 µ g\/mL, add 100 \u0026amp;micro droppers to each sample; L, so that the solution covered the entire sample area and incubated for 20 min at 20-37 ° C. \u003cbr\u003e      Note: Proteinase K can penetrate the cell membrane and nuclear membrane, so that the staining reagents in the subsequent steps can fully enter the nucleus for reaction and improve the labeling efficiency. Too long of the incubation time will increase the risk of the tissue section falling off the carrier plate in the subsequent washing step, and too short of the incubation time may cause inadequate permeability treatment and affect the labeling efficiency. To get a better result, Proteinase K concentration, incubation time and temperature optimization should be carried out according to the different types of tissue samples. \u003cbr\u003e      d. Rinse the sections twice with PBS for 5 min each time, blot off excess liquid with filter paper, and keep the processed sample moist in a wet box. \u003cbr\u003e        Note: this step must be to Proteinase K washing clean, otherwise it will seriously interfere with subsequent tag response. \u003cbr\u003e        e. Turn step \u003cstrong\u003e2. TUNEL \u003c\/strong\u003e\u003cstrong\u003ereaction \u003c\/strong\u003e. \u003c\/p\u003e\n\u003cp\u003e(5) Positive processing (this step is only carried out for positive control, and the other samples are directly carried out for TUNEL reaction step) \u003cbr\u003e        A. 10\u0026amp;times with ddH\u003csub\u003e2\u003c\/sub\u003eO at a ratio of 1:10; DNase I dilute Buffer into 1 \u0026amp; times; DNase I Buffer for later use. \u003cbr\u003e        B. add 100 \u0026amp; micro; L 1× DNase I Buffer to the processed samples, sample cover all areas, balance 5 min at room temperature. \u003cbr\u003e        C. use 1 \u0026amp; times; DNase I Buffer to 1:100 diluted DNase I (2 U \/ \u0026amp; mu; L) to finish working liquid concentration 20 U\/mL. \u003cbr\u003e        D. to Buffer, to join 100 \u0026amp; mu; L DNase I working solution at a concentration of 20 U\/mL was incubated for 10 min at room temperature. \u003cbr\u003e        e. Discard the DNase I working solution and wash twice with PBS. \u003cbr\u003e        f. Turn step \u003cstrong\u003e2. TUNEL \u003c\/strong\u003e\u003cstrong\u003ereaction \u003c\/strong\u003e. \u003cbr\u003e\u003cstrong\u003e2, TUNEL \u003c\/strong\u003e\u003cstrong\u003ereaction \u003cbr\u003e\u003c\/strong\u003e(1) Prepare TUNEL reaction solution (ready to use) : \u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 83.9685%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e   td \u0026gt;\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e5 samples \u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\" samples\u003e td \u0026gt; tr \u0026gt;\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003ethe TdT enzyme  td \u0026gt;\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003cp\u003e1 μ L  p \u0026gt; td \u0026gt;\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\u003cp\u003e5 μ L  p \u0026gt; td \u0026gt;\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\n\u003cp\u003e10 μ \u0026gt;\u003c\/p\u003e\n\u003cp\u003e p\u0026gt;\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e\u003cp\u003eTUNEL Reaction Buffer  p \u0026gt; td \u0026gt;\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\u003cp\u003e49 μ L  p \u0026gt; td \u0026gt;\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\n\u003cp\u003e  245μ \"L\u003c\/p\u003e p\u0026gt;\u003c\/td\u003e\n\u003ctd\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\n\u003cp\u003e490 μ \"\u0026gt;\u003c\/p\u003e\n\u003cp\u003e p\u0026gt;\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 48.1247%; text-align: center;\"\u003e\u003cp\u003eTUNEL reaction liquid volume  p \u0026gt; td \u0026gt;\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.7558%; text-align: center;\"\u003e\n\u003cp\u003e50 μ \"\u0026gt;\u003c\/p\u003e\n\u003cp\u003e p\u0026gt;\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 15.6017%; text-align: center;\"\u003e\n\u003cp\u003e250 μ \"\u0026gt;\u003c\/p\u003e\n\u003cp\u003e p\u0026gt;\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 14.181%; text-align: center;\"\u003e\n\u003cp\u003e500 μ L  p \u0026gt; td \u0026gt; tr \u0026gt; tbody \u0026gt; table \u0026gt;\u003cbr\u003e\u003c\/p\u003e\n\u003cp\u003e(2) smear of adherent cells, cells or tissue section \u003cbr\u003e\u0026amp; have spent     a. Add 50 \u0026amp;mu to each sample; L TUNEL reaction solution so that the reaction solution uniformly covers the sample. The cells were incubated at 37 ° C in the dark for the appropriate time (the recommended staining time for cells was 30min-1h, and the recommended staining time for tissues was 2h). \u003cbr\u003e        Note: 50 μ L TUNEL reaction solution is suitable for smear, section or 96 well plate (other different well plates can appropriately adjust the volume of TUNEL reaction solution and cover the cells). \u003cbr\u003e      If the sample to be tested is a smear, section, or in a 24-well, 12-well, or 6-well plate, you can use an anti-evaporation film, or try to use a zipped bag or other appropriate material to cut yourself into a round plastic sheet slightly smaller than the hole, drop the TUNEL reaction solution and cover the sample, to prevent the TUNEL reaction solution from evaporating, And the TUNEL reaction solution can be uniformly covered the sample. \u003cbr\u003e      b. Discard the TUNEL reaction solution, rinse twice with PBS, and then rinse 3 times with 0.1% Triton X-100 (prepared in PBS containing 5 mg\/mL BSA) for 5 min each time, so that the free unreacted label can be removed relatively cleanly. \u003cbr\u003e      c. (optional) add appropriate concentration of 5\u0026amp;mu to each sample; g\/mL of DAPI staining solution was incubated at room temperature in the dark for 5 min. After staining, DAPI staining solution was discarded and rinsed twice with PBS for 5 min each time. \u003cbr\u003e      d. (optional) Slice sealing: Drop 50 \u0026amp;mu per sample; L Anti-fluorescence quench sealer (anti-fluorescence quench sealer may not work with some dyes, pre-experiment is recommended to test the match before the experiment), cover the cover glass and gently hit the cover glass with the blunt end of the tweezers to remove air bubbles to completely seal the piece. \u003cbr\u003e      e.  Blot off excess liquid with filter paper and add 100\u0026amp;mu to the sample area; L PBS to keep the sample moist and immediately observe under a fluorescence microscope. \u003c\/p\u003e\n\u003cp\u003e(3) for suspended cells or cell suspensions \u003cbr\u003e        a. Add 50\u0026amp;mu to each sample tube; L Cells were gently resuspended in TUNEL reaction solution and incubated at 37 ° C in the dark for 30 to 60 min. Cells were gently resuspended at 15 min intervals using a micropipettor. \u003cbr\u003e        b.  After centrifugation at 2000 rpm for 5min, the TUNEL reaction solution was discarded, the cells were gently resuspended by adding an appropriate amount of 0.1% Triton X-100 (prepared in PBS containing 5 mg\/mL BSA), and washed twice for 5min each time so that the free unreacted marker could be removed relatively cleanly. \u003cbr\u003e      c. Add 100 \u0026amp;mu to each sample tube; L concentration is 5μ g\/mL of DAPI staining solution was incubated at room temperature in the dark for 5 min. \u003cbr\u003e      d. add 400 μ L Cells were resuspended in PBS and immediately examined by flow cytometry or observed under a fluorescence microscope after smear. \u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003cp\u003eWhen stored at-20 ℃, component A should be protected from light to avoid repeated freezing and thawing.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"20T","offer_id":41683112886347,"sku":"abs50058-20T","price":258.0,"currency_code":"USD","in_stock":true},{"title":"50T","offer_id":41683112919115,"sku":"abs50058-50T","price":442.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/2da3ade2a4de45769d6155db89cf28b8.jpg?v=1789740068"},{"product_id":"periodic-acid-schiff-pas-stain-kit","title":"Glycogen PAS Staining Solution Kit (Periodic Acid-Schiff Staining Solution Kit)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eI. Materials to be Prepared: 1. 10% formalin fixative. 2. Distilled water. 3. Ethanol. II. Procedure (for reference only): 1. Conventional fixation, typically using 10% formalin, followed by conventional dehydration and embedding. 2. Dewax paraffin sections and place in distilled water; frozen sections are placed directly in distilled water. 3. Rinse in tap water for 2-3 minutes, then rinse twice in distilled water. 4. Place in an oxidizing agent at room temperature for 5-8 minutes, generally no longer than 10 minutes. 5. Rinse once in tap water, then rinse twice in distilled water. 6. Place the specimen in Schiff Reagent and stain in a dark place at room temperature for 10-20 minutes. 7. Rinse in tap water for 10 minutes. 8. Place the specimen in hematoxylin staining solution and stain the nuclei for 1-2 minutes. 9. Differentiate in acidic differentiation solution for 2-5 seconds. 10. Rinse with tap water for 10-15 minutes, then rinse with double-distilled water until the blue color returns. 11. Dehydrate with conventional ethanol in a stepwise fashion. Transparent with xylene, then mount with neutral gum. \u003cbr\u003e\u003cstrong\u003eIII. Staining results:\u003cbr\u003e\u003c\/strong\u003e\u003ctable style=\"border-collapse: collapse; width: 69.1367%; height: 103px; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 9.91032%; height: 22px;\"\u003ePAS-positive substances\u003c\/td\u003e\n\u003ctd style=\"width: 13.0468%; height: 22px;\"\u003eRed or purple\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 9.91032%; height: 22px;\"\u003eCell nucleus\u003c\/td\u003e\n\u003ctd style=\"width: 13.0468%; height: 22px;\"\u003eBlue\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 9.91032%; height: 22px;\"\u003eCytoplasm\u003c\/td\u003e\n\u003ctd style=\"width: 13.0468%; height: 22px;\"\u003eVarious shades of red\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003eNote: The color depth depends largely on the length of time the sample is exposed to the oxidant solution and Schiff Reagent.\u003cbr\u003eNegative Control (Optional):\u003cbr\u003e1. Dissolve 1g of amylase in 100mL of PBS (pH 5.3) and treat for 30-60 minutes. Then place the sample in the oxidant solution along with the other sections. The result should be negative.\u003cbr\u003e2. (Alternative) Treat a saliva slide (filtered and used) for 30-60 minutes and place the sample in the oxidant solution along with the other sections. The result should be negative.\u003cbr\u003e3. (Alternative) If the photograph is using its own sample, skip the oxidant solution step and place the sample directly in Schiff Reagent. The result should be negative.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eGlycogen staining is a common staining method in pathology. McManus first used the periodic acid-Schiff technique in 1946 to visualize mucin. This method is commonly used to visualize glycogen and other polysaccharides. This stain not only reveals glycogen, but also neutral mucinous substances and certain acidic substances, as well as cartilage, the pituitary gland, mold, fungi, pigments, amyloid, and basement membranes. Oxidant solutions can oxidize 1,2-ethylene glycol groups in carbohydrates and related substances, converting them to dialdehydes. The aldehydes then combine with Schiff's reagent to form a fuchsin compound, producing a purple-red color. Because oxidants can also oxidize other intracellular substances, careful selection of the oxidant concentration and oxidation time is crucial. This ensures that the oxidant groups are oxidized to aldehydes without overoxidation.  \u003cbr\u003e       The characteristics of this glycogen PAS staining solution: it adopts our company's unique formula technology, which greatly enhances the staining effect; it has stable performance and strong specificity; it is simple to operate and only takes about 1 hour. \u003cbr\u003e\u003cstrong\u003eProduct components:\u003c\/strong\u003e\u003ctable style=\"border-collapse: collapse; width: 95%; height: 110px; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.8018%; height: 22px; text-align: center;\"\u003e\u003cstrong\u003eName\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 18.3218%; height: 22px; text-align: center;\"\u003e\u003cstrong\u003e4×50mL\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 17.3618%; height: 22px; text-align: center;\"\u003e\u003cstrong\u003e4×100mL\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px; text-align: center;\"\u003e\u003cstrong\u003eSave\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.8018%; height: 22px; text-align: center;\"\u003eReagent (A) Oxidizing agent\u003c\/td\u003e\n\u003ctd style=\"width: 18.3218%; height: 22px; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 17.3618%; height: 22px; text-align: center;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px; text-align: center;\"\u003e4℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.8018%; height: 22px; text-align: center;\"\u003eReagent (B) Schiff Reagent\u003c\/td\u003e\n\u003ctd style=\"width: 18.3218%; height: 22px; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 17.3618%; height: 22px; text-align: center;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px; text-align: center;\"\u003e4℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.8018%; height: 22px; text-align: center;\"\u003eReagent (C) hematoxylin staining solution\u003c\/td\u003e\n\u003ctd style=\"width: 18.3218%; height: 22px; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 17.3618%; height: 22px; text-align: center;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px; text-align: center;\"\u003eRoom temperature, avoid light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 26.8018%; height: 22px; text-align: center;\"\u003eReagent (D) Acidic Differentiation Solution\u003c\/td\u003e\n\u003ctd style=\"width: 18.3218%; height: 22px; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 17.3618%; height: 22px; text-align: center;\"\u003e100mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px; text-align: center;\"\u003eRoom temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Dewax sections as thoroughly as possible, otherwise the staining effect will be affected. \u003cbr\u003e 2. The oxidizing agent should not be used for too long; the optimal oxidation temperature is 18-22°C. \u003cbr\u003e 3. The oxidizing agent solution and Schiff Reagent should be stored airtight at 4°C, away from sunlight and air during use. Before use, it is best to remove the sections and return them to room temperature 30 minutes in advance, then use them in a dark place away from light. \u003cbr\u003e 4. The acidic differentiation solution should be replaced frequently. The differentiation time should be determined by the thickness of the sections, the type of tissue, and the freshness of the acidic ethanol differentiation solution. In addition, sufficient water rinsing time should be provided after differentiation. \u003cbr\u003e 5. The duration of exposure to the oxidizing agent solution and Schiff Reagent is crucial and should be determined by the thickness of the sections and the type of tissue. \u003cbr\u003e 6. This staining solution is commonly used for staining routine tissue sections. For staining cells or extremely thin sections, it is recommended to purchase the Glycogen PAS Staining Kit (for cells), as it contains lower concentrations of oxidizing agent and hematoxylin solution, which prevents overstaining. 7. Keep the staining time of frozen sections as short as possible. 8. For your safety and health, please wear a lab coat and disposable gloves when operating.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at 2-8℃, away from light. Valid for 6 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50mL×4","offer_id":41683113279563,"sku":"abs90053-50mL×4","price":77.0,"currency_code":"USD","in_stock":true},{"title":"100mL×4","offer_id":41683113312331,"sku":"abs90053-100mL×4","price":137.0,"currency_code":"USD","in_stock":true}]},{"product_id":"periodic-acid-schiff-pas-stain-kit-abs90054","title":"Glycogen PAS Staining Kit (Cell-Specific)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eI. Procedure (for reference only): 1. Fix cell and bone marrow smears with PAS fixative for 10-15 minutes. 2. Rinse the fixative with distilled water and air-dry the sections. 3. Add an oxidizing agent dropwise and oxidize at room temperature for 15-20 minutes. Rinse twice with distilled water, 2 minutes each. 4. Add or immerse in Schiff stain, cover, and stain in a dark place at room temperature for 10-20 minutes. 5. (Optional) Rinse twice with sodium sulfite solution, 2 minutes each. 6. Rinse with running water for 5 minutes (primarily for microscopic observation). 7. Add Mayer's hematoxylin stain dropwise and counterstain for 1-2 minutes. Rinse with water, air-dry, and examine under a microscope. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 9.91032%; height: 22px;\"\u003ePAS-positive substances (glycogen or polysaccharide)\u003c\/td\u003e\n\u003ctd style=\"width: 13.0468%; height: 22px;\"\u003eRed or purple-red\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 9.91032%; height: 22px;\"\u003eCell nucleus\u003c\/td\u003e\n\u003ctd style=\"width: 13.0468%; height: 22px;\"\u003eBlue\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\u003ctd cytoplasm: various shades of red. note: the color depth depends largely on length time sample is exposed to oxidant and schiff stain. negative control add ml aqueous solution treat at for minutes. then place in subsequent experiments. result should be negative. a saliva slide ready use minutes it along with other samples.\u003e\u003c\/td\u003e\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eGlycogen staining is a common staining method in pathology. McManus first used the periodic acid-Schiff technique in 1946 to visualize mucin. This technique is commonly used to visualize glycogen and other polysaccharides. It can not only visualize glycogen, but also neutral mucinous substances and certain acidic substances, as well as cartilage, pituitary glands, molds, fungi, pigments, amyloid substances, basement membranes, and more. Oxidants can oxidize the 1,2-ethylene glycol groups in carbohydrates and related substances, converting them to dialdehydes. The aldehydes then combine with Schiff's reagent to form a fuchsin compound, producing a purple-red color. Because oxidants can also oxidize other intracellular substances, careful selection of the oxidant concentration and oxidation time is crucial. This ensures that the oxidant groups are oxidized to aldehydes without overoxidation. \u003cbr\u003e        Glycogen PAS Staining Solution (for cells only) is characterized by the use of unique formula technology, which greatly enhances the staining effect; stable performance and strong specificity; simple operation, only takes 1 hour; lower concentrations of oxidants and hematoxylin, more suitable for staining cells and ultra-thin tissue sections; no hydrochloric acid and ethanol differentiation steps. \u003cbr\u003eProduct components:\u003ctable style=\"border-collapse: collapse; width: 86.6841%; height: 110px; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.2818%; height: 22px;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 18.0018%; height: 22px;\"\u003e5×20mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.2018%; height: 22px;\"\u003e5×50mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px;\"\u003eSave\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.2818%; height: 22px;\"\u003eReagent (A) PAS fixative\u003c\/td\u003e\n\u003ctd style=\"width: 18.0018%; height: 22px;\"\u003e20mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.2018%; height: 22px;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px;\"\u003eNormal temperature\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.2818%; height: 22px;\"\u003eReagent (B) Oxidizing agent\u003c\/td\u003e\n\u003ctd style=\"width: 18.0018%; height: 22px;\"\u003e20mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.2018%; height: 22px;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 2-8℃, protect from light\u0026lt;\/td\u0026gt;\u0026lt;\/tr\u0026gt;\u0026lt;tr style=\" height:\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 31.2818%; height: 22px;\"\u003eReagent (C) Schiff staining solution\u003c\/td\u003e\n\u003ctd style=\"width: 18.0018%; height: 22px;\"\u003e20mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.2018%; height: 22px;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px;\"\u003e2-8℃, protect from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 31.2818%; height: 22px;\"\u003eReagent (D) Sodium sulfite solution\u003c\/td\u003e\n\u003ctd style=\"width: 18.0018%; height: 20mL\u0026lt;\/td\u0026gt;\u0026lt;td style=\" width: height:\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%; height: 22px;\"\u003eRoom temperature, sealed\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 31.2818%;\"\u003eReagent (E) Mayer's hematoxylin staining solution\u003c\/td\u003e\n\u003ctd style=\"width: 18.0018%;\"\u003e20mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.2018%;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.687%;\"\u003e2-8℃, protect from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. The oxidizing time should not be too long. The optimal oxidation temperature is 18-22°C. 2. Avoid excessive sunlight and air when using oxidizing agents and Schiff stain. It is best to remove the oxidizing agent and Schiff stain 30 minutes before use and allow them to return to room temperature. Use in a dark place away from light. 3. The duration of exposure to the oxidizing agent and Schiff stain is crucial and should be determined based on the thickness of the section and the type of cell or tissue. 4. For routine sections, it is recommended to use glycogen PAS stain, as it contains relatively high concentrations of oxidizing agent and hematoxylin solution. 5. For your safety and well-being, please wear a lab coat and disposable gloves when handling.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at 2-8℃, away from light. Valid for 6 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"20mL×5","offer_id":43224071307339,"sku":"abs90054-20mL×5","price":43.0,"currency_code":"USD","in_stock":true},{"title":"50mL×5","offer_id":41683118981195,"sku":"abs90054-50mL×5","price":87.0,"currency_code":"USD","in_stock":true}]},{"product_id":"ldh-cytotoxicity-assay-kit","title":"Lactate Dehydrogenase (LDH) Cytotoxicity Detection Kit (565nm)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eI. Instruments, consumables, and reagents required: 1. Microplate reader (capable of measuring absorbance at 565 nm) and CO2 incubator. 2. 96-well plates with transparent flat bottoms. 3. Centrifuge (optional). 4. Adjustable pipette and tips. 5. Deionized water. II. Reagent Preparation: Assay Buffer: Ready-to-use; equilibrate to room temperature before use; store at 4°C. Lactic Acid Solution: Ready-to-use; equilibrate to room temperature before use; store at 4°C. MTT Solution: Ready-to-use; keep on ice and protected from light throughout the experiment; aliquot and store at -20°C. PES Solution: Ready-to-use; keep on ice and protected from light throughout the experiment; aliquot and store at -20°C. \u003cbr\u003e\u003cstrong\u003eLDH Positive Control:\u003c\/strong\u003e Ready-to-use; keep on ice and protected from light throughout the experiment; store in aliquots at -20°C. \u003cbr\u003e\u003cstrong\u003eNAD+ Solution:\u003c\/strong\u003e Ready-to-use; keep on ice and protected from light throughout the experiment; store in aliquots at -20°C. \u003cbr\u003e\u003cstrong\u003eTriton X-100 (10%):\u003c\/strong\u003e Ready-to-use; equilibrate to room temperature before use; store at 4°C. \u003cbr\u003e\u003cstrong\u003eLDH Reaction Solution:\u003c\/strong\u003e Prepare 10 mL of LDH reaction solution, sufficient for one 96-well plate, by mixing 3.7 mL Assay Buffer, 1.4 mL MTT Solution, 800 μL NAD+ Solution, 100 μL PES Solution, and 4.0 mL Lactic Acid Solution. Prepare freshly for use. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eIII. Experimental steps:\u003c\/strong\u003e\u003cbr\u003e\u003cstrong\u003e1. Determine whether the drug is suitable for this kit\u003c\/strong\u003e\u003cbr\u003eSome drugs may interfere with the reaction system and are therefore not suitable for this kit. Therefore, we recommend performing an initial pilot experiment to determine whether the target drug you plan to use is suitable for this kit. If the pilot experiment shows that the drug has an inhibitory effect, contact Absin technical support. \u003cbr\u003e(1) Culture the cells for more than 24 hours, centrifuge to remove the particles, and obtain the cell culture supernatant. \u003cbr\u003e(2) Add the cell culture supernatant to a 96-well cell culture plate at 200 μL\/well, with 6 replicates. \u003cbr\u003e(3) Add 20 μL of the target drug to 3 wells, and add 20 μL of Assay Buffer to the other 3 wells as control wells. \u003cbr\u003e(4) Transfer 100 μL of each well to a new 96-well assay plate. \u003cbr\u003e(5) Add 100 μL of LDH Reaction Solution to each well. \u003cbr\u003e(6) Incubate the plate at 37°C for up to 30 min. \u003cbr\u003e(7) Read the absorbance at 565 nm using a microplate reader. \u003cbr\u003e(8) Evaluate the absorbance of the drug-treated wells and the control wells. If the drug-treated wells are significantly smaller than the control wells, the drug is not suitable for this kit. \u003cbr\u003e\u003cstrong\u003e2. Sample Assay\u003c\/strong\u003e\u003cbr\u003e(1) Determine the optimal concentration based on cell size and growth rate. Inoculate 200 μL of cells into a 96-well cell culture plate so that the cell density does not exceed 80-90% at the time of testing. \u003cbr\u003e(2) Aspirate the growth medium from the 96-well plate. Wash the cells once with PBS, then replace the growth medium with low-serum medium containing 1% serum and incubate for another 1 h. \u003cbr\u003e(3) Add 200 μL of 1% low-serum medium (without cells) to three wells as background controls and three wells as LDH positive controls (optional). \u003cbr\u003e(4) Induce cytotoxicity by the desired method and add 20 μL of drug to the appropriate wells in triplicate. Add 20 μL of Assay Buffer to three wells containing cells as spontaneous release, and add 20 μL of Assay Buffer to three wells containing only 1% low-serum medium without cells as background control. \u003cbr\u003e(5) Incubate the plate in a CO\u003csub\u003e2\u003c\/sub\u003e incubator at 37°C for the desired experimental time. \u003cbr\u003e(6) One hour before the scheduled assay, add 20 μL of 10% Triton X-100 solution to three wells containing cells as maximum release, and add 20 μL of LDH Positive Control to three wells containing only 1% low-serum medium without cells as positive control. Mix by pipetting several times and continue incubating in the CO\u003csub\u003e2\u003c\/sub\u003e incubator. \u003cbr\u003e(7) Centrifuge the 96-well tissue culture plate at 400 g for 5 minutes (optional, but recommended). \u003cbr\u003e(8) Transfer 100 μL of cell supernatant to a new 96-well assay plate. \u003cbr\u003e(9) Add 100 μL of LDH Reaction Solution to each well. \u003cbr\u003e(10) Incubate the plate at 37°C for up to 30 minutes. \u003cbr\u003e(11) Read the absorbance at 565 nm using a microplate reader. \u003cbr\u003e(12) Subtract background A565 levels from all wells. \u003cbr\u003e\u003cstrong\u003eNote:\u003c\/strong\u003e The drug concentration added should be the final concentration in the reaction. The results of each experiment are calculated as \"percent cytotoxicity,\" or the percentage of the total amount of LDH contained in the target cells. Therefore, for each experiment, there must be a set of control wells in which all target cells are killed using the 10% Triton X-100 solution provided in the kit. These are the \"maximum release levels.\" Likewise, in every experiment, there must be a set of control wells in which no cytotoxic agent or cytotoxic cells are added, resulting in the lowest (spontaneous) LDH release. These are the \"spontaneous release\" wells. Cells treated with cytotoxic agents will release an amount of LDH that lies between the maximum release level and the spontaneous release level. \u003cbr\u003e\u003cbr\u003e\u003cstrong\u003eIV. Calculation of Results:\u003c\/strong\u003e\u003cbr\u003eThe following formula is used to calculate \"Cytotoxicity Percentage\".\u003cbr\u003eCytotoxicity Percentage (%) = (A \u003csub\u003eSample\u003c\/sub\u003e - A \u003csub\u003eSpontaneous\u003c\/sub\u003e) \/ (A \u003csub\u003eMaximum Release\u003c\/sub\u003e - A \u003csub\u003eSpontaneous\u003c\/sub\u003e) times 100.\u003cbr\u003eA \u003csub\u003eSample\u003c\/sub\u003e, absorbance of the sample treated with toxic reagent at 565 nm;\u003cbr\u003eA \u003csub\u003eSpontaneous\u003c\/sub\u003e, absorbance of spontaneous release of the sample at 565 nm;\u003cbr\u003eA \u003csub\u003eMaximum Release\u003c\/sub\u003e, absorbance of maximum release of the sample at 565 nm.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTheory \u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eLactate dehydrogenase (LDH) is an oxidoreductase that catalyzes the interconversion of pyruvate and lactate, as well as the interconversion of NADH and NAD+. LDH is a stable enzyme present in all cell types and rapidly released into the cell culture medium upon plasma membrane damage. Therefore, LDH is the most widely used marker in cytotoxicity studies. The Cytotoxicity Detection Kit (Lactate Dehydrogenase Method) provides a simple colorimetric assay for studying cytotoxicity. This assay is based on a typical cytotoxicity assay in which target cells are incubated with cytotoxic chemicals or cytotoxic cells (NK cells, cytotoxic T cells) to induce target cell death and LDH release. The supernatant containing LDH is transferred to the wells of a new 96-well assay plate and mixed with the LDH Reaction Solution. In the presence of LDH, NADH reacts with the tetrazolium salt MTT to generate NAD+ and the reduced form of MTT. This reduced form of MTT exhibits a maximum absorption at 565 nm, and the resulting color intensity directly correlates with the number of cells lysed. After incubation at room temperature for 30 min, the absorbance was read at 565 nm using a microplate reader.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eComposition\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\u003ctable style=\"border-collapse: collapse; width: 96.5652%; height: 176px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 30.1252%; height: 22px; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e96T\u003c\/td\u003e\n\u003ctd style=\"width: 16.5654%; height: 22px; text-align: center;\"\u003e480T\u003c\/td\u003e\n\u003ctd style=\"width: 24.8789%; height: 22px; text-align: center;\"\u003eSave\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 30.1252%; height: 22px; text-align: center;\"\u003eAssay Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.5654%; height: 22px; text-align: center;\"\u003e50mL\u003c\/td\u003e\n\u003ctd style=\"width: 24.8789%; height: 22px; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 30.1252%; height: 22px; text-align: center;\"\u003eLactic Acid Solution\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e5mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.5654%; height: 22px; text-align: center;\"\u003e25mL\u003c\/td\u003e\n\u003ctd style=\"width: 24.8789%; height: 22px; text-align: center;\"\u003e4℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 30.1252%; height: 22px; text-align: center;\"\u003eMTT Solution\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e2mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.5654%; height: 22px; text-align: center;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 24.8789%; height: 22px; text-align: center;\"\u003e-20℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 30.1252%; height: 22px; text-align: center;\"\u003ePES Solution\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e120uL\u003c\/td\u003e\n\u003ctd style=\"width: 16.5654%; height: 22px; text-align: center;\"\u003e600uL\u003c\/td\u003e\n\u003ctd style=\"width: 24.8789%; height: 22px; text-align: center;\"\u003eLDH Positive Control\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e120uL\u003c\/td\u003e\n\u003ctd style=\"width: 16.5654%; height: 22px; text-align: center;\"\u003e300uL\u003c\/td\u003e\n\u003ctd style=\"width: 24.8789%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 30.1252%; height: 22px; text-align: center;\"\u003eNAD+ Solution\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e1mL\u003c\/td\u003e\n\u003ctd style=\"width: 16.5654%; height: 22px; text-align: center;\"\u003e5mL\u003c\/td\u003e\n\u003ctd style=\"width: 24.8789%; height: 22px; text-align: center;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 30.1252%; height: 22px; text-align: center;\"\u003eTriton X-100 (10%)\u003c\/td\u003e\n\u003ctd style=\"width: 16.969%; height: 22px; text-align: center;\"\u003e10 mL\u003c\/td\u003e style=\"width: 24.8789%; height: 22px; text-align: center;\"\u0026gt;4℃\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBackground\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eAssessment of cell death or cytotoxicity is often based on the quantification of plasma membrane damage. LDH is a stable enzyme present in all cell types and is rapidly released into the cell culture medium following plasma membrane damage. Therefore, LDH is the most widely used marker in cytotoxicity studies.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Do not mix components from different batches or manufacturers; otherwise, abnormal results may occur.\u003cbr\u003e2. Avoid creating bubbles when mixing or reconstituted components.\u003cbr\u003e3. Change pipette tips frequently to avoid cross-contamination between components.\u003cbr\u003e4. Before beginning the experiment, ensure that all components and equipment are at the appropriate temperature.\u003cbr\u003e5. Maintain good laboratory habits and wear protective equipment such as gloves, lab coat, mask, and goggles before beginning the experiment.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e-20℃, valid for 6 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTest Range\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e10,000-100,0000 cells\/well\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"96T","offer_id":41683113443403,"sku":"abs580274-96T","price":35.0,"currency_code":"USD","in_stock":true},{"title":"480T","offer_id":41683113476171,"sku":"abs580274-480T","price":104.0,"currency_code":"USD","in_stock":true}]},{"product_id":"lactate-dehydrogenase-cytotoxicity-colorimetric-microplate-assay-kit","title":"Lactate Dehydrogenase (LDH) Cytotoxicity Detection Kit (490nm)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eSynonym\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eLDH Cytotoxicity Assay Kit\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eDetection Type: Colorimetric\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe dye in the kit is susceptible to oxidation and deterioration after dissolution. It should be prepared and used immediately. Storage is not recommended. For long-term storage, the dye components of the kit can be purchased separately.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eApplication\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e细胞裂解物，细胞培养基\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eTransport and store at 4°C, shelf life 6 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100T","offer_id":41683113508939,"sku":"abs580275-100T","price":49.0,"currency_code":"USD","in_stock":true},{"title":"500T","offer_id":41683113541707,"sku":"abs580275-500T","price":147.0,"currency_code":"USD","in_stock":true}]},{"product_id":"dna-content-quantitation-assay-cell-cycle","title":"DNA Content Detection Kit (Cell Cycle)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Induce cell apoptosis using an appropriate method. Simultaneously, establish a negative control group and collect cells. 2. Wash cells once with PBS, collect by centrifugation at 1500 rpm for 5 minutes, adjust the cell concentration to 1 × 106\/mL, and prepare 1 mL of single-cell suspension. 3. After centrifugation of the prepared single-cell suspension, remove the supernatant and fix the cells with 500 μL of 70% pre-cooled ethanol for 2 hours to overnight. Store at 4°C. Wash the fixative with PBS before staining. If necessary, filter the cell suspension once through a 200-mesh cell sieve. 4. Add 100 μL of RNase A solution to the cell pellet, resuspend the cells, and incubate at 37°C in a water bath for 30 minutes. 5. Add 400 μL of PI staining solution, mix thoroughly, and incubate at 4°C in the dark for 30 minutes. 6. Analyze red fluorescence at an excitation wavelength of 488 nm. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eThe cell cycle refers to the entire process that a continuously dividing cell undergoes from the end of one mitotic division to the end of the next. During this process, the cell's genetic material replicates and doubles, and at the end of division, it is evenly distributed between the two daughter cells. The cell cycle can be divided into interphase and mitosis. Interphase is often divided into the resting phase (G0), the prophase of DNA synthesis (G1), the DNA synthesis phase (S), and the late phase of DNA synthesis (G2). The entire cycle can be represented as G1 →S →G2 →M. DNA cycle analysis can be used to monitor the status of each phase of the cell cycle, that is, the state of cell proliferation. This method utilizes the ability of intracellular DNA to bind to fluorescent dyes (such as propidium iodide (PI). Different DNA content in cells at different stages of the cycle, resulting in different binding of fluorescent dyes, and different fluorescence intensities detected by flow cytometry. \u003cbr\u003e        During cell apoptosis, cytoplasm and chromatin condense, nuclear lysis occurs, and apoptotic bodies form, resulting in changes in the cells' light scattering properties. In the early stages of apoptosis, the cells' forward-angle light scattering ability decreases significantly, while their 90° angle light scattering ability increases or remains unchanged. In the late stages of apoptosis, both forward-angle and 90° angle light scattering signals decrease. Therefore, apoptotic cells can be observed by flow cytometry by measuring changes in light scattering. When cells are stained with PI, a population of cells with low DNA staining appears before the normal G0\/G1 population due to reduced total DNA content. This sub-diploid peak (sub-G1) appears before the G1 peak, representing the apoptotic population. \u003cbr\u003e          This kit can be used to monitor DNA content (cell cycle) in cultured cells (suspension and adherent). \u003cbr\u003e\u003cstrong\u003eProduct composition:\u003c\/strong\u003e\u003ctable style=\"border-collapse: collapse; width: 95%; height: 66px; margin-left: auto; margin-right: auto;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 23.1218%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003eReagent name\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 14.8%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e50T\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 13.1357%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003e100T\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 17.2966%; text-align: center; height: 22px;\"\u003e\u003cstrong\u003eSave\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 23.1218%; text-align: center; height: 22px;\"\u003eRNase A\u003c\/td\u003e\n\u003ctd style=\"width: 14.8%; text-align: center; height: 22px;\"\u003e5mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.1357%; text-align: center; height: 22px;\"\u003e10mL\u003c\/td\u003e\n\u003ctd style=\"width: 17.2966%; text-align: center; height: 22px;\"\u003e-20℃\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 23.1218%; text-align: center; height: 22px;\"\u003ePI staining solution\u003c\/td\u003e\n\u003ctd style=\"width: 14.8%; text-align: center; height: 22px;\"\u003e20mL\u003c\/td\u003e\n\u003ctd style=\"width: 13.1357%; text-align: center; height: 22px;\"\u003e40mL\u003c\/td\u003e\n\u003ctd style=\"width: 17.2966%; text-align: center; height: 22px;\"\u003e-20℃, protected from light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Protect propidium iodide (PI) staining solution from light during storage and use. \u003cbr\u003e 2. PI is toxic; wear gloves when handling and avoid contamination. \u003cbr\u003e 3. All fluorescent dyes are susceptible to quenching, so it is recommended that testing be completed the same day after staining. \u003cbr\u003e 4. For your safety and well-being, please wear a lab coat and disposable gloves during operation.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e-20℃, avoid light, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50T","offer_id":41683113574475,"sku":"abs90055-50T","price":65.0,"currency_code":"USD","in_stock":true},{"title":"100T","offer_id":41683113607243,"sku":"abs90055-100T","price":97.0,"currency_code":"USD","in_stock":true}]},{"product_id":"mycoplasma-stain-assay-kit","title":"Mycoplasma Stain Assay Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Preparation of cell samples:\u003cbr\u003e(1) For adherent cells, culture the cells in a six-well plate or other multi-well plate or on a coverslip until 50%-80% full. Overfull cell culture can make it difficult to determine whether mycoplasma contamination is present or not.\u003cbr\u003e(2) For suspended cells, centrifuge to precipitate the cells, take a small amount of cells for cell smear, and fully dry them in the air.\u003cbr\u003e2. Hoechst staining solution was diluted 1:10 with PBS (1 volume of Hoechst staining solution was added to 9 volumes of PBS). The diluted Hoechst staining solution should be used within 24 hours.\u003cbr\u003e3. Add an appropriate amount of fixative to fix for 10 to 20 minutes. For one well of a six-well plate, 1 mL of fixative was added. Ensure the fixative adequately covers the sample and never wash the cells prior to fixation. For adherent cells, the culture medium needs to be removed before fixation.\u003cbr\u003e4. Remove the fixing solution and dry it in the air.\u003cbr\u003e5. Add an appropriate amount of Hoechst staining solution diluted 10 times to stain at room temperature for 10 ~ 30min. For one well of a six-well plate, 1 mL of staining solution was added. Ensure that the dyeing solution fully covers the sample, and keep it out of light when dyeing. It can be protected from light with aluminum foil paper.\u003cbr\u003e6. Remove the dyeing solution and dry it in the air.\u003cbr\u003e7. Add the anti-fluorescence quenching sealing solution provided in the kit dropwise, and observe the blue fluorescence under a fluorescence microscope after sealing. Fluorescence microscope observation requires 400 times or 1000 times magnification observation (oil mirror is required). For cell samples without mycoplasma contamination or other prokaryote contamination, only blue fluorescence of the nucleus was observed, and mitochondrial DNA was not stained.\u003cbr\u003eParticulate or filamentous blue fluorescence around the nucleus can be observed in cell samples contaminated with mycoplasma. A large amount of particulate or filamentous blue fluorescence can be observed in cells with heavy mycoplasma contamination.\u003cbr\u003eIn case of bacterial, yeast or mold contamination, although Hoechst staining will also be positive, bacterial, yeast or mold contamination can be observed under an optical microscope, but mycoplasma cannot be observed, so it can be preliminarily distinguished whether it is mycoplasma or other microorganisms.\u003cbr\u003eIf further identification is necessary, further detection can be carried out by inoculating mycoplasma culture and RT-PCR.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eBeilstein\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eAppearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003esolution\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eThe mycoplasma staining detection kit can be used for in situ staining detection of mycoplasma or other prokaryotes in cultured cells. It is mainly used to detect whether there is mycoplasma contamination in cultured cells.\u003cbr\u003eBacterial contamination, yeast contamination or mold contamination in cultured cells are all visible under the light microscope, but mycoplasma contamination is not visible under the light microscope and must be detected by specific detection methods.\u003cbr\u003eThere are many methods to detect mycoplasma contamination, including mycoplasma isolation and culture, mycoplasma-specific enzyme detection, RT-PCR detection and DNA fluorescence staining detection. The kit uses Hoechst staining to detect mycoplasma, which can quickly, effectively and highly sensitively detect mycoplasma contamination.\u003cbr\u003eIf mycoplasma contamination is found, it is recommended to replace non-contaminated cells for culture. The kit can detect at least 100 samples if used for six-well plate sample detection.\u003c\/p\u003e\n\u003cdiv\u003eComponents:\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 50.9855%; height: 84px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.9628%; height: 21px; text-align: center;\"\u003eItem number\u003c\/td\u003e\n\u003ctd style=\"width: 47.3214%; height: 21px; text-align: center;\"\u003eProduct name\u003c\/td\u003e\n\u003ctd style=\"width: 26.3665%; height: 21px; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.9628%; height: 21px; text-align: center;\"\u003eComponent A\u003c\/td\u003e\n\u003ctd style=\"width: 47.3214%; height: 21px; text-align: center;\"\u003eFixative solution\u003c\/td\u003e\n\u003ctd style=\"width: 26.3665%; height: 21px; text-align: center;\"\u003e100ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.9628%; height: 21px; text-align: center;\"\u003eComponent B\u003c\/td\u003e\n\u003ctd style=\"width: 47.3214%; height: 21px; text-align: center;\"\u003eHoechst\u003cspan class=\"font3\"\u003eDyeing solution\u003c\/span\u003e\n\u003c\/td\u003e\n\u003ctd style=\"width: 26.3665%; height: 21px; text-align: center;\"\u003e10ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.9628%; height: 21px; text-align: center;\"\u003eComponent C\u003c\/td\u003e\n\u003ctd style=\"width: 47.3214%; height: 21px; text-align: center;\"\u003eAnti-fluorescence quenching sealing solution\u003c\/td\u003e\n\u003ctd style=\"width: 26.3665%; height: 21px; text-align: center;\"\u003e10ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003ePubChem CID\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e0\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. Hoechst\u003cspan class=\"font3\"\u003eThe dyeing solution is DMSO solution, which will solidify into a solid state at 4 °C. It can be melted in an oven at room temperature or 37 °C before use.\u003c\/span\u003e\u003cbr\u003e2. Hoechst dyeing reagent is harmful to human body, please pay attention to proper protection.\u003cbr\u003e3. The fixative contains glacial acetic acid and has a pungent smell. It should be fixed in the fume hood.\u003cbr\u003e4. Fluorescent dyes all have quenching problems. It is recommended to complete the test on the same day as possible after dyeing.\u003cbr\u003e5. Before detecting mycoplasma, it is best to use antibiotic-free culture medium for 2 to 3 generations, which makes it easier to detect mycoplasma, because some antibiotics can inhibit the growth of mycoplasma.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at 2 ~ 8 ℃ in the dark from light, and the validity period is 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"100T","offer_id":41683113640011,"sku":"abs9252-100T","price":60.0,"currency_code":"USD","in_stock":true}]},{"product_id":"pcr-mycoplasma-test-kit","title":"PCR Mycoplasma Test Kit","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cstrong\u003e1 Sample preparation: \u003c\/strong\u003e Take an appropriate amount of cell culture supernatant to be tested into the clean PCR In the tube, use PCR Instrument 95℃ Heat treatment 5 min After as a template. Serum samples available Mycoplasma Free Water After dilution, take an appropriate amount of sample into a clean PCR In the tube, use PCR Instrument 95℃ Heat treatment 5 min After as a template; \u003cbr\u003e\u003cstrong\u003e2 、 PCR System Preparation: \u003c\/strong\u003e Each experiment needs to set up a negative control (will 1 μL The sample to be tested is replaced with the same amount Mycoplasma Free Water ) vs. positive control (in 1 μL Add to the sample to be tested 0.5 μL Positive Control Later together as Template ), wear disposable masks and gloves during the experiment, and operate carefully to prevent improper operation from introducing exogenous mycoplasma contamination; \u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 54.1393%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 64.1514%; text-align: center;\"\u003e Components \u003c\/td\u003e\n\u003ctd style=\"width: 32.2678%; text-align: center;\"\u003e Volume \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 64.1514%; text-align: center;\"\u003eTemplate\u003c\/td\u003e\n\u003ctd style=\"width: 32.2678%; text-align: center;\"\u003e1ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 64.1514%; text-align: center;\"\u003eMycoplasma Primer Mix\u003c\/td\u003e\n\u003ctd style=\"width: 32.2678%; text-align: center;\"\u003e2ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 64.1514%; text-align: center;\"\u003eMycoplasma PCR Mix （ 2× ） \u003c\/td\u003e\n\u003ctd style=\"width: 32.2678%; text-align: center;\"\u003e10ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 64.1514%; text-align: center;\"\u003eMycoplasma Free Water\u003c\/td\u003e\n\u003ctd style=\"width: 32.2678%; text-align: center;\"\u003eTo 20ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cstrong\u003e3 、 PCR Program settings: \u003c\/strong\u003e\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 73.9816%; height: 152px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 33.5396%; height: 21px; text-align: center;\"\u003e\u003cstrong\u003eStep\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 19.8301%; height: 21px; text-align: center;\"\u003e\u003cstrong\u003eTemperature\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 20.4893%; height: 21px; text-align: center;\"\u003e\u003cstrong\u003eTime\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 19.0233%; height: 21px; text-align: center;\"\u003e\u003cstrong\u003eCycles\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 33.5396%; height: 21px; text-align: center;\"\u003eInitial Denaturation\u003c\/td\u003e\n\u003ctd style=\"width: 19.8301%; height: 21px; text-align: center;\"\u003e98℃\u003c\/td\u003e\n\u003ctd style=\"width: 20.4893%; height: 21px; text-align: center;\"\u003e2 min\u003c\/td\u003e\n\u003ctd style=\"width: 19.0233%; height: 21px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 26px;\"\u003e\n\u003ctd style=\"width: 33.5396%; height: 26px; text-align: center;\"\u003eDenaturation\u003c\/td\u003e\n\u003ctd style=\"width: 19.8301%; height: 26px; text-align: center;\"\u003e98℃\u003c\/td\u003e\n\u003ctd style=\"width: 20.4893%; height: 26px; text-align: center;\"\u003e20 S\u003c\/td\u003e\n\u003ctd style=\"height: 68px; width: 19.0233%; text-align: center;\" rowspan=\"3\"\u003e30\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 33.5396%; height: 21px; text-align: center;\"\u003eAnnealing\u003c\/td\u003e\n\u003ctd style=\"width: 19.8301%; height: 21px; text-align: center;\"\u003e56℃\u003c\/td\u003e\n\u003ctd style=\"width: 20.4893%; height: 21px; text-align: center;\"\u003e25 S\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 33.5396%; height: 21px; text-align: center;\"\u003eExtension\u003c\/td\u003e\n\u003ctd style=\"width: 19.8301%; height: 21px; text-align: center;\"\u003e72℃\u003c\/td\u003e\n\u003ctd style=\"width: 20.4893%; height: 21px; text-align: center;\"\u003e10 S\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 33.5396%; height: 21px; text-align: center;\"\u003eFinal extension\u003c\/td\u003e\n\u003ctd style=\"width: 19.8301%; height: 21px; text-align: center;\"\u003e72℃\u003c\/td\u003e\n\u003ctd style=\"width: 20.4893%; height: 21px; text-align: center;\"\u003e5 min\u003c\/td\u003e\n\u003ctd style=\"width: 19.0233%; height: 21px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 33.5396%; height: 21px; text-align: center;\"\u003eHold\u003c\/td\u003e\n\u003ctd style=\"width: 19.8301%; height: 21px; text-align: center;\"\u003e4-16℃\u003c\/td\u003e\n\u003ctd style=\"width: 20.4893%; height: 21px; text-align: center;\"\u003eForever\u003c\/td\u003e\n\u003ctd style=\"width: 19.0233%; height: 21px; text-align: center;\"\u003e \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cstrong\u003e4 Gel electrophoresis: \u003c\/strong\u003e Take 10 μL PCR Product, using 1% Sepharose gel for electrophoresis detection; \u003cbr\u003e\u003cstrong\u003e5 Results analysis: \u003c\/strong\u003e In each experiment, the mycoplasma contamination and the size of positive band were confirmed by comparing the test results with the negative control and positive control 500 bp Left and right. If there is a band in the negative control test result, it is likely that PCR There is contamination in the system, so it is recommended to re-test to confirm the results. If necessary, also PCR The products were routinely sequenced to determine specific mycoplasma species. \u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003eMycoplasma contamination will cause adverse effects on all aspects of cells, which has become a highly valued problem in cell culture. Therefore, it is necessary to carry out regular detection of mycoplasma contamination. The mycoplasma physical examination test kit is made of PCR Methods, the contamination of mycoplasma in various cultured cell biomaterials (such as cell culture, experimental animal secretions, animal serum, etc.) was specifically detected. \u003cbr\u003eThe mixed primers used in the kit are for Mycoplasma 16s rRNA Specific primers designed for conserved regions of the sequence can be directly used as cell culture fluid PCR Template, specifically amplifying mycoplasma DNA , with matching Mycoplasma PCR Mix （ 2× ) can be completed within one hour. This kit has high detection sensitivity and can detect as low as 20 Copies of mycoplasma. \u003cbr\u003eProduct Components:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 69.5138%; height: 105px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.1118%; height: 21px; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 55.3326%; height: 21px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 19.2063%; height: 21px; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.1118%; height: 21px; text-align: center;\"\u003eA\u003c\/td\u003e\n\u003ctd style=\"width: 55.3326%; height: 21px; text-align: center;\"\u003eMycoplasma PCR Mix （ 2× ）\u003c\/td\u003e\n\u003ctd style=\"width: 19.2063%; height: 21px; text-align: center;\"\u003e1ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.1118%; height: 21px; text-align: center;\"\u003eB\u003c\/td\u003e\n\u003ctd style=\"width: 55.3326%; height: 21px; text-align: center;\"\u003eMycoplasma Primer Mix\u003c\/td\u003e\n\u003ctd style=\"width: 19.2063%; height: 21px; text-align: center;\"\u003e200ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.1118%; height: 21px; text-align: center;\"\u003eC\u003c\/td\u003e\n\u003ctd style=\"width: 55.3326%; height: 21px; text-align: center;\"\u003ePositive Control\u003c\/td\u003e\n\u003ctd style=\"width: 19.2063%; height: 21px; text-align: center;\"\u003e50ul\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 21px;\"\u003e\n\u003ctd style=\"width: 20.1118%; height: 21px; text-align: center;\"\u003eD\u003c\/td\u003e\n\u003ctd style=\"width: 55.3326%; height: 21px; text-align: center;\"\u003eMycoplasma Free Water\u003c\/td\u003e\n\u003ctd style=\"width: 19.2063%; height: 21px; text-align: center;\"\u003e1ml\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e1. This kit can detect at least 11 mycoplasma contamination such as M.orale, M.arginini, M.bovis, M.fermentans, M.gallisepticum, M.hominis, M.pirum, Ureaplasma spp, M.hyorhinis, M.pneumoniae, A.laidlawii, etc.\u003cbr\u003e2. All reagents should be thoroughly thawed and mixed on ice before use, and stored at-20 ℃ after use.\u003cbr\u003e3. Negative control and positive control must be set in each experiment. It is recommended for the experimental group to set a sample template gradient.\u003cbr\u003e4. Masks must be worn during operation, and PCR operating standards must be strictly followed to prevent the introduction of external pollution from affecting the experimental results.\u003cbr\u003e5. In order to ensure the reliability and stability of cell experiments, it is recommended to conduct regular mycoplasma contamination detection.\u003cbr\u003e6. For your safety and health, please wear a lab coat and disposable gloves.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStored at-20 ℃, shelf life is 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50T","offer_id":41683113672779,"sku":"abs9588-50T","price":35.0,"currency_code":"USD","in_stock":true}]},{"product_id":"qpcr-mycoplasma-test-kit","title":"Mycoplasma Detection Kit (qPCR)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\u003ctable\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eUsage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1. Preparation:\u003cbr\u003e1, Reagent Kit Preparation:\u003cbr\u003eKeep the kit components on ice or 4℃, make sure it is fully dissolved before starting the operation.\u003cbr\u003e2, Consumables and equipment required:\u003cbr\u003eFluorescence quantification PCRInstrument, vortex instrument, centrifuge\u003cbr\u003eHigh-precision pipettes and disposable sterile, enzyme-free, low-adsorption filter tips (1-2.5 μL、0.5-10 μL、10-100 μL、20-200 μL、100-1000 μL) 5 mLSterile enzyme-free low-adsorption centrifuge tubes\u003cbr\u003eSterile enzyme-free eight-tube strip or 96hole qPCRplate\u003cbr\u003e2. Operation process:\u003cbr\u003e1Detailed operation steps:\u003cbr\u003e（1) Extraction of test samples:\u003cbr\u003eRecommended use“Host cell residues DNA(Magnetic beads method) Sample pretreatment kit (abs60544)”Extract samples DNAWhen adding the sample, add 4μLof Internal ControlWhen extracting samples, it is recommended to extract them synchronously DNAThe dilution served as a negative control.\u003cbr\u003e（2) qPCRPreparation of reaction solution:\u003cbr\u003e   ①According to the number of reference materials and samples to be tested (usually 2-3Calculate the number of wells required for the reaction:\u003cbr\u003e       Number of reaction wells=（1No-template control NTC+1Positive control PC+Sample to be tested) ×Number of multiple holes\u003cbr\u003e   ②Calculate the required amount based on the number of reaction wells qPCR MIXTotal amount:\u003cbr\u003e       qPCR MIX =(Number of reaction wells+2or3)×12.5μL（2×qPCR Reaction MIX)+(Number of reaction wells +2or 3)×2.1μL(Mycoplasma Primer\u0026amp;Probe MIX)+(Number of reaction wells+2or3)×0.4μL（Internal Control)( 2or 3is the operating loss)\u003cbr\u003e③Dissolve all reagents on ice, shake gently to mix, centrifuge briefly, and prepare as shown in the following table:\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 51.9007%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 29.4854%; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 13.6092%; text-align: center;\"\u003eSingle reaction volume\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 29.4854%; text-align: center;\"\u003e2×qPCR Reaction MIX\u003c\/td\u003e\n\u003ctd style=\"width: 13.6092%; text-align: center;\"\u003e 12.5 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 29.4854%; text-align: center;\"\u003eMycoplasma Primer\u0026amp;Probe MIX\u003c\/td\u003e\n\u003ctd style=\"width: 13.6092%; text-align: center;\"\u003e 2.1 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 29.4854%; text-align: center;\"\u003eInternal Control\u003c\/td\u003e\n\u003ctd style=\"width: 13.6092%; text-align: center;\"\u003e 0.4 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 29.4854%; text-align: center;\"\u003eTotal volume\u003c\/td\u003e\n\u003ctd style=\"width: 13.6092%; text-align: center;\"\u003e 15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp\u003e【Note】:If it is added during extractionIC, then prepareqPCR MIXWhen equal volume DNAdiluent instead IC.\u003cbr\u003e④will be prepared qPCR MIX, gently shake to mix, centrifuge briefly, press15 μL\/Hole divided into eight tubes or 96In the orifice plate.\u003cbr\u003e⑤Will DNADiluent press 10 μL\/The hole is added to the sub-packagingqPCR MIXEight-tube or96In the well plate, see the table below for specific sample additions.\u003cbr\u003eMelted and unused DNAThe diluent can be 2-8If not used for a long time, please store at-20℃.\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 74.3109%;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 26.5891%; text-align: center;\"\u003ePositive PC\u003c\/td\u003e\n\u003ctd style=\"width: 44.1514%; text-align: center;\"\u003ePCeach 10 μL+Required qPCR MIXquantity 15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 26.5891%; text-align: center;\"\u003eNo template control NTC\u003c\/td\u003e\n\u003ctd style=\"width: 44.1514%; text-align: center;\"\u003eDNADilution 10 μL+Required qPCR MIXquantity 15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 26.5891%; text-align: center;\"\u003eSamples to be tested\u003c\/td\u003e\n\u003ctd style=\"width: 44.1514%; text-align: center;\"\u003eThe samples to be tested 10 μL+Required qPCR MIXquantity 15 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cbr\u003e（3) qPCRAdd sample:\u003cbr\u003e①Place the required reagents on ice, shake gently to mix, centrifuge briefly, and add the sample (total volume 25 μL):\u003cbr\u003e②The experiment can use sterile enzyme-free eight-tube or 96When performing the reaction in the well plate, it is necessary to remove the bubbles in the reaction system and centrifuge to the bottom of the tube to prepare for the reaction.\u003cbr\u003e（4) qPCRReaction program parameter settings:\u003cbr\u003eThe United States ABI 7500 Real-Time PCR System, software version 2.06For example.\u003cbr\u003eA,exist Experimental PropertiesCreate a blank new program on the page as shown below:\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/c1d4cf92fa4b46bfb4604e192438c7cb.png\" alt=\"\" width=\"501\" height=\"249\"\u003e\u003cbr\u003e①Experiment name: Modify the experiment name.②Experiment type: Select a program (Quantitation-Standard Curve).③Reagents: Select the experimental method (TaqMan® Reagents).\u003cbr\u003eB, create the product detection probe, selectselect\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/e538885129294a459a44bfca2eec6610.png\" alt=\"\" width=\"149\" height=\"27\"\u003e ①interface, click\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/bd59680c758d4f18babcd31ef94f1c95.png\" alt=\"\" width=\"87\" height=\"23\"\u003e ②Create a detection probe and an internal reference probe. TargetNamemiddleEdit the probe name and name it.Reporter③andQuencher④Select the desired fluorophore and quencherThis product detects fluorescent groups.FAM, the quenching group isNone, selection of internal reference fluorescent groupsCY5, the quenching group isNone, the reference fluorescence isROX.pointhit\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/de3d02cf33b044e98cccca75bc6ae140.png\" alt=\"\" width=\"90\" height=\"23\"\u003e⑤Create samples, in the corresponding Sample name⑥Name the sample in the column.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/e1e493279eee4393a075994f0fea1aec.png\" alt=\"\" width=\"499\" height=\"221\"\u003e\u003cbr\u003eC,existPlate Setup \u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/71f88632f4ac40068aeba9ceb63446ff.png\" alt=\"\" width=\"140\" height=\"19\"\u003e①Interface, place the positive control wells at Task②onecolumnset upPlacefor “S”③, the no-template control NTCKong ZaiTask②One column is set to“N”④, place the sample hole to be tested inTask②One column is set to “U”⑤, and in the corresponding Sample nameName the sample in a column (Figure LegendonlyforGinsengTest).\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/dae0a928687f44eca672d8dc2c5f060c.png\" alt=\"\" width=\"504\" height=\"251\"\u003e\u003cbr\u003eD, set up the reaction program:\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/1b06279652a4424d9f35b2675e873f50.png\" alt=\"\" width=\"490\" height=\"201\"\u003e\u003cbr\u003e【Note】Some devices do not allow the fluorescence collection step to be set to 30sor shorter; use ABI7000、 ABI 7300and ABI 7500At least 31sFor other equipment, please consult the instrument manufacturer.\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/982e8de482634f62934f407d94e87dfc.png\" alt=\"\" width=\"491\" height=\"300\"\u003e\u003cbr\u003eE, click RunInterface \"Start Run\"Button to proceedPCRDetermination.\u003cbr\u003e3. Result Analysis\u003cbr\u003eThe United States ABI 7500 Real-Time PCR System, software version 2.06For example.\u003cbr\u003e1,exist Analysisof Amplification Plot①In the panel, clickReanalyse②, initially check whether the shape of the amplification curve is normal. AnalysisofViewWell Table③In the panel, CtOne column can read the sample to be tested CtDetection value. (The illustration is for reference only)\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/a82d2799a5cb44e497d0085828ea9cac.png\" alt=\"\" width=\"488\" height=\"223\"\u003e\u003cbr\u003e2The parameter settings for result analysis need to be based on the specific model and software version used, and can generally be automatically interpreted by the instrument.\u003cbr\u003e3、NTC,Positive PCThe results must meet\u003cbr\u003e\u003cimg src=\"https:\/\/absin.oss-cn-shanghai.aliyuncs.com\/absin-china\/20240801\/767c8361a1694c509fa9a6f94e437aea.png\" alt=\"\" width=\"469\" height=\"297\"\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eDescription\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eMycoplasmas are prokaryotes that commonly contaminate cells in the laboratory. The main sources of contamination include cells and virus strains, as well as animal-derived raw materials such as serum, chicken embryos, and primary cells, and the operators themselves. According to statistics, approximately 15%-35% of cells are contaminated with mycoplasmas. Mycoplasmas can alter a range of host cell characteristics, including structure and function, leading to inaccurate experimental results. They can also interfere with cell metabolism and growth, alter nucleic acid synthesis, affect cell antigenicity, cause chromosomal changes, interfere with viral replication, and mimic viral functions. \u003cbr\u003e This test kit offers high specificity and sensitivity, and can be used to qualitatively detect mycoplasma contamination in master cell banks, working cell banks, viral seed batches, control cells, and cells used in clinical treatment. It can be used in conjunction with the Host Cell Residual DNA (Magnetic Bead Method) Sample Pretreatment Kit (abs60544). Detection Principle: According to \u0026lt;2.6.7\u0026gt; of the European Pharmacopoeia, nucleic acid amplification (NAT) can replace culture and indicator cell culture methods after appropriate validation. Fluorescent probe (TaqMan®) qPCR is characterized by high sensitivity and specificity, making TaqMan® the preferred method for mycoplasma detection. TaqMan® fluorescent probes are oligonucleotide probes with a fluorescent group (reporter) at the 5' end, such as FAM, CY5, or VIC, and a quencher at the 3' end, such as TAMRA or BHQ1. During PCR amplification, a specific fluorescent probe is added along with a pair of primers. When the probe is intact, the fluorescent signal emitted by the fluorescent group is absorbed by the quencher. During PCR amplification, the 5'-3' exonuclease activity of the Taq enzyme cleaves and degrades the probe, separating the fluorophore and quencher. This allows the fluorescence monitoring system (fluorescence quantitative PCR instrument) to detect a fluorescent signal. This means that each time a DNA strand is amplified, a fluorescent molecule is formed, ensuring that the accumulation of the fluorescent signal is completely synchronized with the formation of PCR products. This kit utilizes a multiplex PCR approach, using two different fluorescent probes to detect the target sequence and the internal reference, respectively. \u003cbr\u003eProduct components:\u003cbr\u003e\u003ctable style=\"border-collapse: collapse; width: 83.6162%; height: 132px;\" border=\"1\"\u003e\u003ctbody\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.41183%; height: 22px; text-align: center;\"\u003eNumber\u003c\/td\u003e\n\u003ctd style=\"width: 30.8096%; height: 22px; text-align: center;\"\u003eName\u003c\/td\u003e\n\u003ctd style=\"width: 13.0525%; height: 22px; text-align: center;\"\u003eSpecifications\u003c\/td\u003e\n\u003ctd style=\"width: 23.736%; height: 22px; text-align: center;\"\u003eStorage conditions\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.41183%; height: 22px; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 30.8096%; height: 22px; text-align: center;\"\u003eMycoplasma Primer\u0026amp;Probe MIX\u003c\/td\u003e\n\u003ctd style=\"width: 13.0525%; height: 22px; text-align: center;\"\u003e120 µL\u003c\/td\u003e\n\u003ctd style=\"width: 23.736%; height: 22px; text-align: center;\"\u003e-20℃ and below light protection\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.41183%; height: 22px; text-align: center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"width: 30.8096%; height: 22px; text-align: center;\"\u003e2×qPCR Reaction MIX\u003c\/td\u003e\n\u003ctd style=\"width: 13.0525%; height: 22px; text-align: center;\"\u003e700 \u0026amp; micro; height: 22px; text-align: center;\"\u0026gt;3\u003c\/td\u003e\n\u003ctd style=\"width: 30.8096%; height: 22px; text-align: center;\"\u003eInternal Control\u003c\/td\u003e\n\u003ctd style=\"width: 13.0525%; height: 22px; text-align: center;\"\u003e220 µL\u003c\/td\u003e\n\u003ctd style=\"width: 23.736%; height: 22px; text-align: center;\"\u003e-20℃ and below\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 22px;\"\u003e\n\u003ctd style=\"width: 8.41183%; height: 22px; text-align: center;\"\u003e4\u003c\/td\u003e\n\u003ctd style=\"width: 30.8096%; height: 22px; text-align: center;\"\u003ePositive Control\u003c\/td\u003e\n\u003ctd style=\"width: 13.0525%; height: 22px; text-align: center;\"\u003e500 \u0026amp; micro; center;\"\u0026gt;5\u003c\/td\u003e\n\u003ctd style=\"width: 30.8096%; height: 22px; text-align: center;\"\u003eDNA Dilution 1.5 mL × 3 -20°C or below\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eTechnical Index\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eLimit of Detection: 100% detection rate for 10 standard strains of Mycoplasma species listed in various pharmacopoeias, including Mycoplasma oralis, Mycoplasma hyorhinis, and Mycoplasma pneumoniae. Limit of Blank: No detection was detected in 24 replicates of DNA diluted directly. Precision: Ten replicates of the same Mycoplasma-positive reference sample yielded positive results, with a coefficient of variation of Ct values of \u0026lt;5%. Specificity: No detection was detected when 300 pg\/μL genomic DNA from common biological products (e.g., Human, Vero, CHO, HEK293, E. coli, Pichia pastoris) was used as a sample. No detection was detected when DNA from other engineered bacteria (e.g., Lactobacillus acidophilus and Lactobacillus fermentum) was used as a sample. No detection was detected when DNA from negative matrices such as PBS and 10% fetal bovine serum was used as a sample. \u003cbr\u003eDurability: \u003cbr\u003eMycoplasma standard strains can be stably detected under different dilution matrices (such as DMEM medium, CHO serum-free medium, etc.), product freeze-thaw, product transportation, and multiple instrument model changes (such as ABI 7500, Bio-Rad CFX Opus96, Roche LightCycler 96, etc.).\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eGeneral Notes\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003e(1) This kit has passed the stability verification (freeze-thaw and other factors) and does not require repackaging. \u003cbr\u003e (2) The preparation and loading environment of negative samples (2×qPCR Reaction MIX, Primer \u0026amp; Probe MIX and DNA diluent, etc.) and positive samples (PC and test samples, etc.) must be separated into different areas and cannot be operated in the same area. The preparation personnel must dress neatly, wear masks, gloves and clean clothes. \u003cbr\u003e (3) Pay attention to changing the pipette tips in time between different loading steps to avoid cross contamination, avoid opening the lid for a long time, and avoid liquid sticking to the wall when using a pipette. \u003cbr\u003e (4) The kit must be used within the validity period. It is not recommended to mix related reagents from different batches. \u003cbr\u003e (5) It is recommended that all components in the kit be thawed in a low temperature environment before use, and it must be ensured that they are fully dissolved. Each component must be fully mixed before use to ensure the homogeneity of the reagents. The mixed reagents must be briefly centrifuged so that all the liquid on the tube wall and lid is concentrated at the bottom of the tube. If precipitates are found in the DNA diluent, it is recommended to incubate at 37°C to completely dissolve the DNA diluent. \u003cbr\u003e (6) Only by strictly following the operating instructions and using all the reagents provided with this kit can the best detection results be guaranteed. \u003cbr\u003e (7) The final test results are closely related to the effectiveness of the reagents, the operator's operating methods, and the test environment.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\u003cstrong\u003eStorage Temp.\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd\u003eStore at -20℃, valid for 12 months.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\u003c\/div\u003e","brand":"Absin","offers":[{"title":"50T","offer_id":41683113771083,"sku":"abs9925-50T","price":968.0,"currency_code":"USD","in_stock":true}]}],"url":"https:\/\/www.antbioinc.com\/collections\/cell-biology-kit.oembed?page=5","provider":"AntBio","version":"1.0","type":"link"}