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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.

Key Product Details

Differentiated Mouse CD4+ Cells Secrete IL-17. 
(2)
Intracellular Cytokine Staining of Differentiated Mouse Th17 Cells. 
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CDK017
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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

Citations for CellXVivo Mouse Th17 Cell Differentiation Kit (5)

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Customer Reviews (7)

4.9 out of 5
7 customer ratings
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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 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 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: Jim Hsiao
    Verified Customer | Posted 10/26/2017
    CellXVivo Mouse Th17 Cell Differentiation Kit CDK017

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FAQs

  • How long are the cytokines stable after they are diluted in the Mouse Th17 Differentiation Media, in Catalog # CDK017? Can I prepare the Differentiation Media with cytokines on day 1 of the protocol, and use the same media on day 3, or do I need to add freshly diluted cytokines at each refresh step?

    We have not performed any stability testing to detemine how long diluted cytokines are stable in media without any activity loss. However, during the development of the CDK017 kit, we prepared complete Mouse Th17 Differentiation Media with cytokines added on day 1 and used it until the end of the experiment (refreshing media on day 3 and harvesting on day 5). Cytokines are most stable in medias containing 5-10% serum. We recommend X-VIVO 15 Chemically Defined, Serum-free Hematopoietic Cell Medium (Lonza, or equivalent), which contains HSA that will help stabilize the cytokines for shorter periods.

  • Can I sterile filter the Mouse Th17 Differentiation Media in Catalog # CDK017 after the addition of the cytokines?

    Yes, the complete media may be filtered after addition of cytokines. A 0.2 micron low protein binding sterile filter may be used.

Product Documents

Certificate of Analysis

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