The RayBio® Human TBNK Flow Cytometry Kit is designed to determine the percentages 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+).
TBNK Flow Cytometry Assay Kit
TBNK Flow Cytometry Assay Kit
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Product Description
Specifications
| Size | 50 tests, 100 tests |
| Shipping Type | Blue ice |
| Storage | 4°C |
Description
The RayBio® Human TBNK Flow Cytometry Kit is designed to determine the percentages 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
- 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 or similar
- De-ionized (DI) H2O
- Centrifuge
- Vortex mixer
- Optional: Fluorescent Counting Beads
Antibody Information
| Antibody | Clone | Isotype |
|---|---|---|
| RayBright® Violet 450 Anti-human CD19 | 4G7 | Mouse, IgG1 κ |
| RayBright® Violet 500 Anti-human CD8 | OKT8 | Mouse, IgG2a κ |
| RayBright® Blue 488 Anti-human CD3 | OKT3 | Mouse, IgG2a κ |
| PE Anti-human CD4 | OKT4 | Mouse, IgG2b κ |
| PerCP Anti-human CD45 | 9.4 | Mouse, IgG2a |
| APC Anti-human CD16 | 3G8 | Mouse, IgG1 κ |
| APC Anti-human CD56 | 5.1H11 | Mouse, IgG1 κ |
Assay Procedure
- Dilute the RayBio® 10X Whole Blood Lysing Solution 10-fold with de-ionized (DI) H2O to prepare 1X lysing solution.
- Prepare the antibody cocktail in a 1.5 mL tube. For each test, add 30 μL of the Antibody Diluent and 2.5 μL of each antibody reagent. The final volume of antibody cocktail should be 50 μL per test.
- Label each 12 × 75-mm tube. Add 50 μL of well-mixed anticoagulated whole blood from each sample to the bottom of the appropriate tube.
- Add 50 μL of antibody cocktail to the bottom of each tube and vortex gently to mix.
- Incubate for 20 minutes at room temperature, protected from light.
- Add 1 mL of 1X lysing solution to each tube and vortex gently to mix.
- Incubate for 20 minutes at room temperature (20°C–25°C), protected from light.
- Centrifuge the samples at 500 x g for 5 minutes. Carefully discard the supernatant without disturbing the cell pellet.
- Resuspend the pellet in 0.5 mL of 1X lysing solution. Samples are 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.