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CellXVivo Mouse Th17 Cell Differentiation Kit

For the differentiation of Th17 cells from a preparation of CD4+ T cells isolated from mouse splenocytes.
Differentiated Mouse CD4+ Cells Secrete IL-17. 
(2)
Intracellular Cytokine Staining of Differentiated Mouse Th17 Cells. 
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CDK017

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 Th17 phenotype with reagents provided in the CellXVivo Mouse Th17 Cell Differentiation Kit using the following procedure:

  • Isolate CD4+ T cells from mouse splenocytes
  • Culture CD4+ T cells in reagents provided in kit
  • Verify Th17 cell expansion after 5 days in culture
 

 

Reagents Provided

Reagents Supplied in the CellXVivo Mouse Th17 Cell Differentiation Kit (Catalog # CDK017):

  • Hamster Anti-Mouse CD3
  • Rat Anti-Mouse CD28
  • Mouse Th17 Reagent 1
  • Mouse Th17 Reagent 2
  • Mouse Th17 Reagent 3
  • Mouse Th17 Reagent 4
  • Mouse Th17 Reagent 5
  • Reconstitution Buffer 1
  • Reconstitution Buffer 2
  • Reconstitution Buffer 3
  • 20X Wash Buffer

 

Other Supplies Required

Reagents

  • Laboratory mice
  • XVIVO™ 15 Chemically Defined, Serum-free Hematopoietic Cell Medium (Lonza, or equivalent)
  • MagCellect™ Mouse Naïve CD4+ T Cell Isolation Kit (R&D Systems, Catalog # MAGH115, 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

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

Coat the desired tissue culture plate or flask with Hamster Anti-Mouse CD3 antibody.

Coat the desired tissue culture plate or flask

Isolate mouse splenocytes.

Isolate mouse splenocytes

Perform a cell count.

Perform a cell count

Perform a cell count.

Perform a cell count

Suspend 1 x 106 naïve CD4+ T cells/mL in Mouse Th17 Differentiation Media. Culture the cells on plates or flask pre-coated with Hamster Anti-Mouse CD3 antibody.

Suspend cells in media

Refresh with Mouse Th17 Differentiation Media on day 3. Culture for 2 more days.

Refresh with Mouse Th17 Differentiation Media

Verify Th17 cell differentiation by analyzing cytokine expression via flow cytometry or ELISA (optional).

Verify Treg cell differentiation

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Customer Reviews (7)

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  • CellXVivo Mouse Th17 Cell Differentiation Kit
    Name: Anonymous
    Verified Customer | Posted 01/28/2021
    Many thanks to this kit! The paper is accepted with the help of this kit!
    CellXVivo Mouse Th17 Cell Differentiation Kit Serum CDK017
  • CellXVivo Mouse Th17 Cell Differentiation Kit
    Name: Anonymous
    Verified Customer | Posted 03/09/2020
    I was struggling with Th17 differentiation for a couple of months. Finally figure it out with this FANTASTIC kit!. I highly recommend it!
    CellXVivo Mouse Th17 Cell Differentiation Kit Serum CDK017
  • CellXVivo Mouse Th17 Cell Differentiation Kit
    Name: Anonymous
    Verified Customer | Posted 10/09/2018
    This kit has made my assay much more easier. It's an all-in-one!
  • CellXVivo Mouse Th17 Cell Differentiation Kit
    Name: Anonymous
    Verified Customer | Posted 04/05/2018
  • CellXVivo Mouse Th17 Cell Differentiation Kit
    Name: Anonymous
    Verified Customer | Posted 04/03/2018
  • CellXVivo Mouse Th17 Cell Differentiation Kit
    Name: Anonymous
    Verified Customer | Posted 04/02/2018
  • CellXVivo Mouse Th17 Cell Differentiation Kit
    Name: Anonymous
    Verified Customer | Posted 10/26/2017
    CellXVivo Mouse Th17 Cell Differentiation Kit Serum CDK017

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Product Datasheets

Certificate of Analysis

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