{"product_id":"annexin-v-apc-cyanine7-7-aad-apoptosis-kit","title":"Annexin V-APC\/Cyanine7\/7-AAD Apoptosis Kit","description":"\u003ch2\u003eAnnexin V-APC\/Cyanine7\/7-AAD Apoptosis Kit\u003c\/h2\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eIntroduction\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003eElabscience® Annexin V-APC\/Cyanine7\/7-AAD Apoptosis Kit is developed to detect the apoptosis of suspension cells and adherent cells.\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003eAnnexin V is a member of the annexin family, which binds to phosphatidylserine (PS) in a calcium-dependent manner. Annexin V-APC\/Cyanine7, the APC\/Cyanine7-conjugated format, binds specifically to the PS on the outer leaflet of apoptotic cell membrane and can be detected with flow cytometry or fluorescence microscopy.\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003eDue to the loss of integrity of membrane, 7-AAD can enter late apoptotic or necrotic cells to stain DNA. Cells at different apoptotic stages can be distinguished by using Annexin V and 7-AAD.\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003e\u003cem\u003eJurkat cells were treated with 5 μM Camptothecin and detected with this kit.\u003c\/em\u003e\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003eJurkat cells were cultured with (Left) or without (Right) 5 μM Camptothecin for 4 h. Annexin V-APC\/Cyanine7 single-positive cells were early apoptotic cells, Annexin V-APC\/Cyanine7 and 7-AAD double-positive cells were necrotic or late apoptotic cells, and 7-AAD single-positive cells were naked nuclei.\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\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\u003eAPC\/Cyanine7\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eEx\/Em (nm)\u003c\/td\u003e\n\u003ctd\u003e650;760\/780\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eChannel set\u003c\/td\u003e\n\u003ctd\u003eAPC\/Cy7\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"30%\"\u003eOther reagents required\u003c\/td\u003e\n\u003ctd\u003ePBS(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 class=\"MsoNormal\"\u003e\u003cstrong\u003eReagent Preparation\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003e1×Annexin V Binding Buffer: Dilute Annexin V Binding Buffer (10×) with deionized water to 1×Annexin V Binding Buffer before use.\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003e \u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003e\u003cstrong\u003eStaining Procedure\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\u003c\/ol\u003e\n\u003cp\u003eControl settings for Annexin V experiments are critical to experimental results. It is strongly recommended to read this article before the experiment: https:\/\/www.elabscience.com\/resource-cell_function-detail-984\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eOne-step process\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\u003c\/ol\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 to resuspend the cells.\u003c\/li\u003e\n\u003cli\u003eAdd 5 µL of Annexin V-APC\/Cyanine7 and 5 µL of 7-AAD to each tube.\u003c\/li\u003e\n\u003cli\u003eGently vortex the cells and incubate at room temperature for 15~20 min in the dark.\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.\u003cbr\u003eNote: Annexin V-APC\/Cyanine7 can be detected in APC\/Cy7 channel while 7-AAD can be detected in \u003cspan style=\"font-size: 0.875rem;\"\u003ePerCP\/Cy5.5 channel.\u003c\/span\u003e\n\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eTwo-step process\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\u003c\/ol\u003e\n\u003col\u003e\u003c\/ol\u003e\n\u003col\u003e\u003c\/ol\u003e\n\u003col\u003e\u003c\/ol\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 100 μL of 1×Annexin V Binding Buffer to resuspend the cells.\u003c\/li\u003e\n\u003cli\u003eAdd 2.5 µL of Annexin V-APC\/Cyanine7 and 2.5 µL of 7-AAD to each tube\u003cbr\u003e(Attributed to the higher resolution of two-step protocol, half the amount of the reagents can still guarantee a result of matched quality as in the one-step protocol. It's also recommended that users titrate the reagents for optimal performance in specific models.)\u003c\/li\u003e\n\u003cli\u003eGently vortex the cells and incubate at room temperature for 15~20 min in the dark.\u003c\/li\u003e\n\u003cli\u003eAdd 400 μL of 1×Annexin V Binding Buffer to the tube, and mix gently.\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.\u003cbr\u003eNote: Annexin V-APC\/Cyanine7 can be detected in APC\/Cy7 channel while 7-AAD can be detected in \u003cspan style=\"font-size: 0.875rem;\"\u003ePerCP\/Cy5.5 channel.\u003c\/span\u003e\n\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. 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":"200 Assays","offer_id":40441639731258,"sku":"E-CK-A230-200","price":444.95,"currency_code":"USD","in_stock":true},{"title":"100 Assays","offer_id":40441639764026,"sku":"E-CK-A230-100","price":292.95,"currency_code":"USD","in_stock":true},{"title":"50 Assays","offer_id":40441639796794,"sku":"E-CK-A230-50","price":168.95,"currency_code":"USD","in_stock":true},{"title":"20 Assays","offer_id":40441639829562,"sku":"E-CK-A230-20","price":84.95,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0722\/7785\/files\/annexin-v-apc-cyanine7-7-aad-apoptosis-kit-200-assays-elabscience-biotechnology-cell-assays-21922883895354.jpg?v=1752500782","url":"https:\/\/www.msesupplies.com\/en-gb\/products\/annexin-v-apc-cyanine7-7-aad-apoptosis-kit","provider":"MSE Supplies","version":"1.0","type":"link"}