Annexin V Binding Buffer (10×)

Annexin V Binding Buffer (10×)

Introduction

Elabscience® Annexin V Binding Buffer (10×) is developed to identify apoptotic and necrotic cells.

Product Details

Properties

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

1. . 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.
2. 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.
3. 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.
4. Gently vortex the cells and incubate at room temperature for 15~20 min in the dark.
5. 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

1. This kit is for research use only.
2. For your safety and health, please wear the lab coat and disposable gloves before the experiments.