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Assay Procedure
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- Take 100?μL peripheral blood anticoagulated by EDTA and add to the bottom of 5?mLtube,
- Add 20?μL labeled antibody to the bottom of flow tube mixing with the whole blood, incubate for 20?minutes at room temperature away from light,
- Add 2 ml1×RBC lysis buffer, incubate for 10?minutes away from light after mixing, dissolve red blood cells (recommended: RBC lysing Solution 10×,Cat.: FXP001),
- Sample tube is set to 1000 rpm centrifugation for 5?minutes, discard the supernatant,
- Add 2?mLPBS wash buffer to resuspend the cells, then1000 rpm centrifugation for 5?minutes, discard the supernatant,
- Add 0.5?mLPBS wash buffer to resuspend the cells and detect by flow cytometry (sample should be determined on the day on the machine and can also be added fixation overnight at 4?°C then measured).
- [PBS wash buffer: PBS +1?% FBS +0.1?% NaN3, Cat.: FXP005]
- [Cell fixation: 2?% formaldehyde solution]
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