Sample preparation experiments for apoptosis detection by flow cytometry

Summary

This experimental method was obtained from the official website of the Fourth Military Medical University

Operation method

Sample preparation experiments for indirect immunofluorescence labeling by flow cytometry

Move

Apoptosis was induced according to the experimental protocol, single-cell suspensions were prepared, and apoptosis was detected by specific binding of Annexin V antibody to phosphatidylserine (PS) using the Annexin V-FITC Apoptosis Detection Kit.

1. 10× of binding buffer was diluted to 1× with distilled water and ice incubated.

2. Suspend the cells in a cold environment and wash the cells twice with PBS (centrifugation at 800-1000rpm for 5min).

3. Discard the supernatant and resuspend the cells by adding 490 μl of pre-cooled binding buffer (cell concentration of 105～106/ml).

4. Add 5μl Annexin V-FITC and 5μl PI to the cell suspension and mix gently.

5. Place the tube on ice and incubate for 10 minutes away from light.

6. Perform the FCM assay.

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