CellXVivo Human Th2 Cell Differentiation Kit Best Seller
R&D Systems, part of Bio-Techne | Catalog # CDK002
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, human CD4+ T cells can be differentiated into Th2 cells using the following procedure:
- Coat a plate with Mouse Anti-Human CD3 Antibody
- Isolate human naïve CD4+ T cells from a PBMC preparation
- Culture naïve CD4+ T cells in Human Th2 Differentiation Media for 13 days
- Refresh the Human Th2 Differentiation Media every 3-4 days
- Analyze culture media and cells for a Th2-specific cytokine production profile
Reagents Supplied in the CellXVivo™ Human Th2 Cell Differentiation Kit (Catalog # CDK002):
- Mouse Anti-Human CD3 Antibody
- Human Th2 Reagents
- Reconstitution Buffers
- Wash Buffer (20X)
Reagents
- MagCellect™ Human Naïve CD4+ T Cell Isolation Kit (Catalog # MAGH115 or equivalent)
- Monensin
- PMA
- Ficoll-Hypaque™
- X-VIVO™ 15 Chemically Defined, Serum-free Hematopoietic Cell Medium (Lonza, or equivalent)
- Sterile deionized water
- Ionomycin
- Penicillin-Streptomycin
Equipment
- Tissue culture flasks and/or plates
- Hemocytometer
- 37 °C and 5% CO2 humidified incubator
- Centrifuge
- Pipettes and pipette tips
Coat wells of a 24-well plate with Mouse Anti-Human CD3 Antibody.
Isolate PBMCs from human blood (e.g., using Ficoll-Hypaque™ density centrifugation).
Isolate human naïve CD4+ T cells from PBMCs (e.g., using magnetic cell selection).
Perform a cell count.
Suspend 1-2 x 105 naïve CD4+ T cells/mL in Human Th2 Differentiation Media.
Culture the cells on plates pre-coated with CD3 Antibody for 13 days.
Refresh the Diferentiation Media every 3-4 days.
Stimulate the cells with mitogens.
Verify Th2 cell differentiation by analyzing cytokine expression using flow cytometry.
The Th2 cells are now ready for further downstream applications.