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CellXVivo Human Th1 Cell Differentiation Kit

R&D Systems, part of Bio-Techne | Catalog # CDK001

For the differentiation of Th1 cells from a preparation of naïve CD4+ T cells.
R&D Systems, part of Bio-Techne

Key Product Details

Verification of Th1 Cell Identity using Flow Cytometry. 
(3)
Discontinued Product
CDK001 has been discontinued. View all Th1 Cell Kits products.

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, human CD4+ T cells can be differentiated into Th1 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 Th1 Differentiation Media for 5 days
  • Stimulate cells with monensin
  • Analyze culture media and cells for a Th1-specific cytokine production profile
 

 

Reagents Provided

Reagents Supplied in the CellXVivo™ Human Th1 Cell Differentiation Kit (Catalog # CDK001):

  • Mouse Anti-Human CD3 Antibody
  • Human Th1 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™
  • RPMI 1640
  • L-Glutamine
  • Ionomycin
  • Penicillin
  • Streptomycin
  • Fetal Bovine Serum (FBS)
  • beta-Mercaptoethanol

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 R&D Systems MagCellect™ Human Naïve CD4+ T Cell Isolation Kit).

Isolate T Cells from PBMCs

Perform a cell count.

Perform a cell count

Suspend 1-2 x 105 naïve CD4+ T cells/mL in media supplemented with Human Th1 Reagent 1 and Human Th1 Reagent 2.
Culture the cells on plates pre-coated with Mouse Anti-Human CD3 Antibody for 5 days.

Refresh the Diferentiation Media every 3-4 days.

Suspend cells in media

Stimulate the cells with mitogens.

Stimulate the cells with mitogens

Verify Th1 cell differentiaiton by analying cytokine expression via flow cytometry.

Verify Th1 cell differentiation
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Product Documents for CellXVivo Human Th1 Cell Differentiation Kit

Certificate of Analysis

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