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CellXVivo Human Th2 Cell Differentiation Kit Best Seller

R&D Systems, part of Bio-Techne | Catalog # CDK002

For the differentiation of Th2 cells from a preparation of naïve CD4+ T cells.
R&D Systems, part of Bio-Techne

Key Product Details

Detection of IL-5, IFN-gamma, and IL-17 in Differentiated Human Th2 Cells. 
(3)
Discontinued Product
CDK002 has been discontinued. View all Th2 Cell Kits products.

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 Provided

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)

 

Other Supplies Required

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

 

Procedure Overview

Coat wells of a 24-well plate with Mouse Anti-Human CD3 Antibody.

Coat wells of a 24-well plate

Isolate PBMCs from human blood (e.g., using Ficoll-Hypaque™ density centrifugation).

Isolate PBMCs from human blood

Isolate human naïve CD4+ T cells from PBMCs (e.g., using magnetic cell selection).

Isolate T Cells from PBMCs

Perform a cell count.

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.

Suspend cells in media

Stimulate the cells with mitogens.

Re-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.

Verify Th2 cell differentiation
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Product Documents for CellXVivo Human Th2 Cell Differentiation Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.