{"product_id":"annexin-v-egfp-reagent","title":"Annexin V-EGFP Reagent","description":"\u003ch2\u003eAnnexin V-EGFP Reagent\u003c\/h2\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eIntroduction\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003eElabscience® Annexin V-EGFP Reagent is developed to identify apoptotic cells.\u003c\/p\u003e\n\u003cp\u003eAnnexin V is a member of the annexin family, which binds to phosphatidylserine (PS) in a calcium-dependent manner. Annexin V-EGFP, the EGFP-conjugated format, binds specifically to the PS on the outer leaflet apoptotic cell membrane and can be detected with flow cytometry or fluorescence microscopy.\u003c\/p\u003e\n\u003cp\u003eCells at different apoptotic stages can be distinguished by using Annexin V and membrane impermeable DNA dyes like Propidium Iodide (PI), 7-Amino Actinomycin D (7-AAD) or 4',6-Diamidino-2-Phenylindole (DAPI).\u003c\/p\u003e\n\u003cp\u003e\u003cem\u003eJurkat cells were treated with 5 μM Camptothecin and detected with this kit.\u003c\/em\u003e\u003c\/p\u003e\n\u003cp\u003eJurkat cells were cultured with (Left) or without (Right) 5 μM Camptothecin for 4 h. Annexin V-EGFP single-positive cells were early apoptotic cells, Annexin V-EGFP and PI double-positive cells were necrotic or late apoptotic cells, and PI single-positive cells were naked nuclei.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003ch3\u003e\u003cb\u003eProduct Details\u003c\/b\u003e\u003c\/h3\u003e\n\u003cp\u003e\u003cb\u003eProperties\u003c\/b\u003e\u003c\/p\u003e\n\u003ctable class=\"dtl_table2\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eApplication\u003c\/td\u003e\n\u003ctd\u003eAnnexin V\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eDetection method\u003c\/td\u003e\n\u003ctd\u003eFluorometric Method\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eSample type\u003c\/td\u003e\n\u003ctd\u003eCell samples\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eAssay time\u003c\/td\u003e\n\u003ctd\u003e40min\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eDetection instrument\u003c\/td\u003e\n\u003ctd\u003eFlow Cytometer\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eDye Type\u003c\/td\u003e\n\u003ctd\u003eEGFP\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eEx\/Em (nm)\u003c\/td\u003e\n\u003ctd\u003e488\/510\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eChannel set\u003c\/td\u003e\n\u003ctd\u003eFITC\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eOther reagents required\u003c\/td\u003e\n\u003ctd\u003eAnnexin V Binding Buffer(E-CK-A151), PBS(E-BC-R187), Deionized water\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eStorage\u003c\/td\u003e\n\u003ctd\u003e2~8°C, shading light\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eShipping\u003c\/td\u003e\n\u003ctd\u003eIce bag\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eExpiration date\u003c\/td\u003e\n\u003ctd\u003e12 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eStaining Procedure\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli\u003eInduce apoptosis of suspension cells with reagents of interest. Collect cell cultures, centrifuge at 300 ×g for 5 min and discard the supernatant. Add PBS to wash the cells and resuspend the cells gently followed by the cell counting.\u003c\/li\u003e\n\u003cli\u003eSplit the cell suspension into tubes, 1~5×105 cells for each, centrifuge at 300 ×g for 5 min and discard the supernatant. Add PBS to wash the cells and discard the supernatant. Add 500 μL of 1 × Annexin V Binding Buffer [E-CK-A151] to resuspend the cells.\u003c\/li\u003e\n\u003cli\u003eAdd 5 µL of Annexin V-EGFP Reagent and 5 µL of DNA dye (PI [E-CK-A161], 7-AAD [E-CK-A162]) or DAPI [E-CK-A163]) to each tube.\u003cbr\u003e\n\u003c\/li\u003e\n\u003cli\u003eGently vortex the cells and incubate at room temperature for 15~20 min in the dark.\u003cbr\u003e\n\u003c\/li\u003e\n\u003cli\u003eAnalyze the cells immediately with proper machine settings. Otherwise, place the cells on ice in the dark and analyze within 1 h.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eStorage\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003eStore at 2~8°C for one year in the dark. Avoid freeze \/ thaw cycles.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eNotes\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli\u003eThis kit is for research use only.\u003c\/li\u003e\n\u003cli\u003eWhen detecting adherent cells, the suspension cells generated after induction of apoptosis should be collected and detected together with the subsequently collected adherent cells.\u003c\/li\u003e\n\u003cli\u003eMechanical damage caused by digestion of adherent cells should be avoided as much as possible. At the same time, trypsin digestion solution should not contain EDTA as much as possible, because EDTA will affect the binding of Annexin V to phosphatidylserine.\u003c\/li\u003e\n\u003cli\u003eIf trypsin containing EDTA is used, cells should be washed thoroughly after harvesting to ensure that EDTA is removed.\u003c\/li\u003e\n\u003cli\u003eDetect apoptosis as soon as possible after staining to avoid the increase number of apoptosis or necrosis.\u003c\/li\u003e\n\u003cli\u003eAvoid extended exposure of the samples to direct light to protect the fluorophores from quenching.\u003c\/li\u003e\n\u003cli\u003eFor your safety and health, please wear the lab coat and disposable gloves before the experiments.\u003c\/li\u003e\n\u003c\/ol\u003e","brand":"Elabscience Biotechnology","offers":[{"title":"500 Tests","offer_id":40538080149562,"sku":"E-CK-A119-500","price":803.95,"currency_code":"USD","in_stock":true},{"title":"200 Tests","offer_id":40538080182330,"sku":"E-CK-A119-200","price":399.95,"currency_code":"USD","in_stock":true},{"title":"100 Tests","offer_id":40538080215098,"sku":"E-CK-A119-100","price":278.95,"currency_code":"USD","in_stock":true},{"title":"50 Tests","offer_id":40538080247866,"sku":"E-CK-A119-50","price":189.95,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0722\/7785\/files\/E-CK-A119.jpg?v=1719825108","url":"https:\/\/www.msesupplies.com\/en-gb\/products\/annexin-v-egfp-reagent","provider":"MSE Supplies","version":"1.0","type":"link"}