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

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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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