Flow cytometry (FACS) is an immunophenotyping technique that analyze the expression of cell surface and intracellular molecules. Flow cytometry rapidly measures the specific characteristics of a large number of individual cells. Living cells in suspension are stained with specific, fluorescently labeled antibodies and then analyzed with a flow cytometer. The intensity of fluorescence detected by cytometer is directly proportional to the antigen density or the characteristics of the cell being measured.


Reagents Instruments and Consumables
Washing Buffer Eppendoff
Primary Antibody Fluorochrome-labeled Secondary Antibody
Centrifuge Flow Cytometer


  1. Harvest, wash the cells and then determine the total cell number.
  2. Resuspend the cells to approximately 1-5 x 106 cells/mL in PBS.
  3. Add 100 μl of cell suspension to each tube.
  4. Add 0.1-10 μg/mL of the primary antibody binding cells antigen. Dilutions, if necessary, should be made in PBS.
  5. Incubate for 60 min at 4°C.
  6. Wash the cells 3 times by centrifugation at 400g for 5 min and resuspend them in ice cold PBS.
  7. Dilute the fluorochrome-labeled secondary antibody in 3% BSA/PBS at the optimal dilution and add to cell suspension.
  8. Incubate for 60 min at 4°C in dark.
  9. Wash the cells 3 times by centrifugation at 400g for 5 min and resuspend them in PBS.
  10. Analyze the cells on the flow cytometer. It is recommended that analysis is performed on the same day.
For research use only, not directly for clinical use.
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