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CellXVivo Mouse Th1 Cell Differentiation Kit

For the differentiation of Th1 cells from a preparation of CD4+ T cells isolated from mouse splenocytes.
Th1-differentiated Mouse CD4+ Cells Secrete High Levels of IFN-?.
(3)
Upregulation of T-bet, TNF-alpha, and IL-18R alpha in Th1-polarized Mouse CD4 Cells. 
Intracellular Cytokine Staining of Differentiated Mouse Th1 Cells. 
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CDK018

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, CD4+ T cells isolated from mouse splenocytes are differentiated ex vivo into the Th1 phenotype with reagents provided in this kit and using the following procedure:

  • Isolate CD4+ T cells from mouse splenocytes
  • Culture CD4+ T cells in reagents provided in kit
  • Verify Th1 cell expansion after 6 days in culture
 

 

Reagents Provided

This kit contains the following reagents for the ex vivo differentiation of mouse Th1 cells.

  • Hamster Anti-Mouse CD3, Th1
  • Mouse Th1 Reagent 1
  • Mouse Th1 Reagent 2
  • Mouse Th1 Reagent 3
  • Mouse Th1 Reagent 4
  • Reconstitution Buffer 1
  • Reconstitution Buffer 2
  • 20X Wash Buffer

 

Other Supplies Required

Reagents

  • Laboratory mice
  • X-VIVO™ 15 Chemically Defined, Serum-free Hematopoietic Cell Medium (Lonza, or equivalent)
  • MagCellect Mouse Naïve CD4+ T Cell Isolation Kit (R&D Systems, Catalog # MAGM205, or equivalent)
  • Penicillin/Streptomycin (optional)
  • Cell Activation Cocktail 500X (Tocris®, Catalog # 5476)

Equipment

  • Tissue culture plates and/or flasks
  • Sterile deionized water
  • Pipettes and pipette tips
  • Inverted microscope
  • Hemocytometer
  • 37 °C, 5% CO2 incubator
  • Centrifuge

 

Procedure Overview

Protocol for mouse Th1 cell differentiation using the CellXVivo Mouse Th1 Cell Differentiation Kit.

Note: Results may vary due to strain, age, and/or the health of the mice used for isolation.

Coat the desired tissue culture plate with Hamster Anti-Mouse CD3,
Th1 antibody.

Coat the desired tissue culture plate

Isolate mouse splenocytes.

Isolate mouse splenocytes

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

Perform a cell count

Perform a cell count.

Perform a cell count

Suspend 1 x 106 naïve CD4+ T cells/mL in Mouse Th1 Differentiation Media. Culture the cells on plates pre-coated with Hamster Anti-Mouse CD3 antibody.

Suspend cells in media

Harvest cells on day 3.

Dilute cells 1:10 with fresh Mouse Th1 Differentiation Media.

Culture cells in a new flask for an additional 3 days.

Harvest cells on Day 3

Verify Th1 cell differentiation by analyzing cytokine expression via flow cytometry or ELISA (optional).

Verify Th2 cell differentiation

Citations

Currently there are no citations for CellXVivo Mouse Th1 Cell Differentiation Kit.

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    Verified Customer | Posted 04/05/2018
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Product Datasheets

Certificate of Analysis

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