{"title":"Apoptosis Detection 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":"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":"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":"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":"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":"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"}],"url":"https:\/\/www.antbioinc.com\/collections\/apoptosis-detection-kit.oembed?page=2","provider":"AntBio","version":"1.0","type":"link"}