{"product_id":"bmms-ua090055","title":"Cytokine Kit for Inducing Osteoclast Differentiation of Mouse Bone Marrow Cells (BMMs)","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\n\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\n\u003ctable style=\"width: 100%; height: auto;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eSpecies\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eMouse\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003ePhysical Appearance\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eLyophilized powder\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReconstitution\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003eReconstitute at 0.1-1 mg\/ml according to the size in ultrapure water after rapid centrifugation.\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cspan\u003e\u003cbr\u003e·12 months from date of receipt, lyophilized powder stored at -20 to -80℃.\u003cbr\u003e·3 months, -20 to -80℃ under sterile conditions after reconstitution.\u003cbr\u003e·1 week, 2 to 8℃ under sterile conditions after reconstitution.\u003cbr\u003e·Please avoid repeated freeze-thaw cycles.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003ch4\u003eBackground\u003c\/h4\u003e\n\u003cdiv\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e\n\u003ch4\u003eComponents\u003c\/h4\u003e\n\u003cdiv\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: auto;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003eComponent\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003eS Size\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003eM Size\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003eL\u003c\/span\u003e\u003cspan style=\"color: #000000;\"\u003e Size\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eM-CSF Protein, Mouse\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003e5\u003c\/span\u003e\u003cspan style=\"color: #000000;\"\u003e0μg\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003e1\u003c\/span\u003e\u003cspan style=\"color: #000000;\"\u003e0\u003c\/span\u003e\u003cspan style=\"color: #000000;\"\u003e0μg\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan\u003eRANK L\/TNFSF11 \u003c\/span\u003e\u003cspan\u003eProtein, Mouse\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003e10μg\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003e5\u003c\/span\u003e\u003cspan style=\"color: #000000;\"\u003e0μg\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003ctd style=\"text-align: center; vertical-align: middle; border-top: 1px solid #000000; border-right: 1px solid #000000; border-bottom: 1px solid #000000;\"\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cspan style=\"color: #000000;\"\u003e1\u003c\/span\u003e\u003cspan style=\"color: #000000;\"\u003e0\u003c\/span\u003e\u003cspan style=\"color: #000000;\"\u003e0μg\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cbr\u003e\n\u003cp\u003e\u003cspan\u003e1ml system\u003c\/span\u003e\u003cspan\u003e：\u003c\/span\u003e\u003cspan\u003eM-CSF need \u003c\/span\u003e\u003cspan\u003e180ng in all , RANKL total require \u003c\/span\u003e\u003cspan\u003e150ng\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e\n\u003ch4\u003eProtocol\u003c\/h4\u003e\n\u003cdiv\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.Bone Marrow Cell Seeding\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.1After red blood cell lysis of freshly isolated mouse bone marrow cells, resuspend the cells in α-MEM complete medium containing 10% fetal bovine serum (FBS) and perform cell counting.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.2Dilute the cell density to 2 × 10⁵ cells\/mL, add M-CSF to a final concentration of 30 ng\/mL, and seed the cells into 24-well cell culture plates pre-loaded with sterile coverslips, with 1 mL of cell suspension added to each well.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e1.3 Incubate the plates statically in a cell incubator at 37°C with 5% CO₂ for 5 days, and change the medium (containing 30 ng\/mL M-CSF) every 2 days.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2.Osteoclast Induction and Differentiation\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2.1Aspirate and discard the old medium in the wells. Add complete medium containing 30 ng\/mL M-CSF to the negative wells, and add complete medium containing 30 ng\/mL M-CSF and 50 ng\/mL RANKL protein to the positive wells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e2.2Thereafter, change the medium every 2 days. Add complete medium containing 30 ng\/mL M-CSF to the negative wells, and add complete medium containing 30 ng\/mL M-CSF and 50 ng\/mL RANKL protein to the positive wells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3.Morphological Observation and Identification of Cells\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3.1 Morphological observation: Regularly observe the changes in cell morphology under an inverted microscope starting from the induction. Typically, osteoclast-like structures can be observed 4 days after RANKL induction.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003e3.2 TRAP staining verification: Perform tartrate-resistant acid phosphatase (TRAP) staining to specifically identify osteoclasts 5–7 days after RANKL induction. Conduct the staining strictly in accordance with the instructions of the TRAP staining kit.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eObserve the cells under a light microscope after staining. The cytoplasm of mature osteoclasts appears red or reddish-purple.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003c\/div\u003e","brand":"UA BIOSCIENCE","offers":[{"title":"S","offer_id":42302185209931,"sku":null,"price":260.0,"currency_code":"USD","in_stock":true},{"title":"M","offer_id":42302185242699,"sku":null,"price":700.0,"currency_code":"USD","in_stock":true},{"title":"L","offer_id":42302185275467,"sku":null,"price":1260.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/AntBioImage_7ee88cf1-1d97-44b8-b9c9-b6bd4b3ba69a.png?v=1776073678","url":"https:\/\/www.antbioinc.com\/products\/bmms-ua090055","provider":"AntBio","version":"1.0","type":"link"}