Annexin V Binding Buffer (10×) Introduction Elabscience® Annexin V Binding Buffer (10×) is developed to identify apoptotic and necrotic cells. Product Details Properties Application Annexin V Storage 2~8°C Shipping Ice bag Expiration date 24 months Instructions Annexin V Binding Buffer (10×) is a 10×concentrated solution. Dilute with DI water to 1×working solution before use. For example: Take 1 mL Annexin V Binding Buffer (10×), dilute with DI water to 10 mL. Experimental Procedure . Induce 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 the cells and resuspend the cells gently followed by the cell counting. Split 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. Add 5 µL of Annexin V- fluorescein and 5 µL of DNA dye (PI[E-CK-A161] or 7-AAD[E-CK-A162] or DAPI[ECK-A163]) to each tube. Gently vortex the cells and incubate at room temperature for 15~20 min in the dark. Analyze the cells immediately with proper machine settings. Otherwise, place the cells on ice in the dark and analyze within 1 h. Storage Store at 2~8°C. Valid for 12 months. Notes This kit is for research use only. For your safety and health, please wear the lab coat and disposable gloves before the experiments.
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