Basic T Cell and B Cell Flow Cytometry Panel

Immunophenotyping of T Cells and B Cells by Flow Cytometry

T cells and B cells are critical mediators of adaptive immunity. This multicolor flow cytometry panel allows for easy identification of T cells and B cells.



Flow Cytometry Panel for Identification of T Cells and B Cells

Marker Clone Fluorochrome Catalog #
CD3 UCHT1* Alexa Fluor® 405 FAB100V
CD19 4G7-2E3 PE FAB4867P
CD45 2D1 Alexa Fluor® 647 FAB1430R

*Designate clones independently validated by HLDA.
Alexa Fluor® is registered trademark of Molecular Probes, Inc.

This multicolor flow cytometry panel was validated on human peripheral blood mononuclear cells (PBMCs).


Flow Cytometry Gating Strategy for TB Cell Basic Panel

Pseudocolor flow cytometry plot showing gating strategy for CD19 B Cells

Multicolor flow cytometry panel to identify human B Cells.. PBMCs were stained with Anti-Human CD45 Alexa Fluor® 647, CD3 Alexa Fluor 405®, and CD19 PE. B cells indicated by CD19+ cells in histogram plot. All antibodies are validated for flow cytometry.



Staining Protocol for TB Cell Basic Panel

Other Supplies Required

  1. Wash human PBMCs (1 x 106 cells per sample) with 2 mL of Staining Buffer (1X) (Catalog # FC001) or other BSA-containing buffer, by spinning at 300 x g for 5 minutes, using 5 mL flow cytometry tubes. Decant/aspirate supernatant.
  2. Fc-block cells with blocking IgG (1 μg IgG/106 cells) for 10 minutes at room temperature.
  3. Add previously titrated amount of each primary conjugated antibody. Vortex tubes.

    Recommended Antibody Concentrations  
    Marker Fluorochrome Recommended Concentration
    CD3 Alexa Fluor® 405 5 μL/106 cells
    CD19 PE 10 μL/106 cells
    CD45 Alexa Fluor® 647 0.25-1 μg/106 cells
  4. (Optional) To a separate tube, add 5 μL of each of the isotype control antibodies. Vortex tubes.
  5. Incubate the mixtures for 30-45 minutes at room temperature in the dark.
  6. At the end of the incubation, wash with 2 mL of Staining Buffer (1X), by spinning at 300 x g for 5 minutes. Decant/aspirate supernatant.