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CellXVivo Mouse Treg Cell Differentiation Kit

R&D Systems, part of Bio-Techne | Catalog # CDK007

For the differentiation of mouse regulatory T cells (Tregs) from a preparation of naïve CD4+ T cells.
R&D Systems, part of Bio-Techne

Key Product Details

Differentiation of CD4+ T Cells into Treg Cells Confirmed by FoxP3 and CD25 Expression. 
(2)

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, naïve mouse CD4+ T cells can be differentiated into Tregs using the following procedure:

  • Coat a plate with Hamster Anti-Mouse CD3 Antibody
  • Isolate naïve CD4+ T cells from mouse splenocytes
  • Culture naïve CD4+ T cells in Mouse Treg Differentiation Media for 5 days
  • Verify differentiation into Treg cells by flow cytometry
 

 

Reagents Provided

Reagents Supplied in the CellXVivo™ Mouse Treg Cell Differentiation Kit (Catalog # CDK007):

  • Hamster Anti-Mouse CD3 Antibody
  • Mouse Treg Reagent 1
  • Mouse Treg Reagent 2
  • Mouse Treg Reagent 3
  • Reconstitution Buffer 1
  • Reconstitution Buffer 2
  • Wash Buffer (20X)

 

Other Supplies Required

Reagents

  • MagCellect™ Mouse Naïve CD4+ T Cell Isolation Kit (R&D Systems, Catalog # MAGH205, or equivalent).
  • RPMI 1640
  • Fetal Bovine Serum (FBS)
  • β-Mercaptoethanol (2-ME)
  • L-Glutamine–Penicillin–Streptomycin solution

Equipment

  • Tissue culture flasks and/or plates
  • Sterile deionized water
  • Microscope
  • Hemocytometer
  • 37 °C and 5% CO2 incubator
  • Centrifuge

 

Procedure Overview

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

Coat wells of a 24-well plate

Prepare a single cell suspension of mouse splenocytes.

Prepare a single cell suspension

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 0.5 - 1 x 106 naïve CD4+ T cells/mL in Mouse Treg Differentiation Media.

Culture the cells on plates pre-coated with CD3 antibody for 5 days.

Suspend cells in media

Verify Treg cell differentiation by analyzing marker expression using flow cytometry.

Mature Treg cells are now ready for further downstream applications.

Verify Treg cell differentiation
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Product Documents for CellXVivo Mouse Treg Cell Differentiation Kit

Certificate of Analysis

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