The RayBio® Human TBNK Flow Cytometry Kit is designed to determine the percentages and absolute counts of mature human lymphocyte subsets in peripheral whole blood (WB). The subsets include T lymphocytes (CD3+), Helper T lymphocytes (CD3+CD4+), Cytotoxic T lymphocytes (CD3+CD8+), B lymphocytes (CD19+) and Natural killer (NK) lymphocytes (CD3-CD16&56+).
The RayBio® Human TBNK Flow Cytometry Kit is designed to determine the percentages and absolute counts of mature human lymphocyte subsets in peripheral whole blood (WB). The subsets include T lymphocytes (CD3+), Helper T lymphocytes (CD3+CD4+), Cytotoxic T lymphocytes (CD3+CD8+), B lymphocytes (CD19+) and Natural killer (NK) lymphocytes (CD3-CD16&56+). Our kit contains individually packed fluorochrome-labeled antibodies, antibody diluent, fluorescent counting beads for absolute number counts (cells/μL) of gated cell populations, and lysing solution to minimize the red blood cells. This kit is easy and convenient to use, and a perfect tool for immunophenotyping peripheral lymphocytes of human whole blood.
Kit Contents
RayBright® Violet 450 Anti-human CD19
RayBright® Violet 500 Anti-human CD8
RayBright® Blue 488 Anti-human CD3
PE Anti-human CD4
PerCP Anti-human CD45
APC Anti-human CD16
APC Anti-human CD56
Fluorescent Counting Beads
Antibody Diluent
RayBio® 10X Whole Blood Lysing Solution
Additional Materials Required
Flow cytometer with violet, blue and red lasers
1.5 mL polypropylene microcentrifuge tubes or similar
12 × 75-mm tubes
De-ionized (DI) H2O
Centrifuge
Vortex mixer
Assay Procedure
Dilute the RayBio® 10X RBCs Lysing Buffer 10-fold with de-ionized (DI) H2O to prepare 1X lysing solution.
Prepare antibody cocktail in a 1.5 mL tube by adding 2.5 μL of the antibody reagents one by one and 32.5 μl of the Antibody Diluent in the kit per single test. The final volume of antibody cocktail per single test is 50 μL.
Label each 12 × 75-mm tube and add 50 μL of well-mixed, anticoagulated whole blood from each sample to the bottom of the tube.
Pipette 50 μL of antibody cocktail from step 2 above into the bottom of each tube and vortex gently to mix.
Incubate for 20 minutes in the dark at room temperature (20°C–25°C).
Thoroughly vortex the Fluorescent Counting Beads in the kit and add exactly 20 μL of beads to each sample tube.
Add 1 mL of 1X lysing solution from step 1 to each tube and vortex gently to mix.
Incubate for 20 minutes in the dark at room temperature (20°C-25°C).
Centrifuge the samples at 500 x g for 5 minutes and discard the supernatant without disturbing the pellet.
Add 0.5 mL of 1X lysing solution from step 1 to each tube. The sample is now ready to be analyzed on a flow cytometer.
Storage/Stability
Store kit at 4°C, protected from light for up to three months for full functionality.
