Annexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. PS is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and PS translocates to the external leaflet. Fluorochrome-labeled Annexin V can then be used to specifically target and identify apoptotic cells. Annexin V Binding Buffer is recommended for use with Annexin V staining.
Products List：Annexin V-PE、7-AAD Viability Staining solution、Annexin V Binding Buffer、Apoptosis Positive Control Solution。
Each lot of this reagent is quality control tested by immunofluorescent staining with flow cytometric analysis (The amount of the reagent is suggested to be used 5 µl/10^5 cells). Since applications vary, the appropriate dilutions must be determined for individual use.
Suggested Staining Protocol：
1. Harvest cell (about 1×106cells per test) then wash with cold PBS.
2. Suspend cells in 1 mL 1× Binding Buffer, 300×g centrifugation for 10 minutes, then remove the Binding Buffer from the cell pellet.
3. Resuspend cells in 1 mL 1× Binding Buffer ,adjust cell concentration to 1×106cells/mL.
4. Add 100 μL of cells (1×105cells) to each labeled tube.
5. Add 5 μL of Annexin V-PE to appropriate tubes.
6. Gently vortex each tube and incubate for 10 minutes in room temperature, protected from light.
7. for 5min in room temperature, protected from light.
8. Add PBS to 500μL and vortex gently.
9. Analyze by flow cytometry in 1 hour.
Keep as concentrated solution. Store at 4°C and protected from prolonged exposure to light. Do not freeze.The validity period is 6 months.
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