CellXVivo Mouse Treg Cell Differentiation Kit

For the differentiation of mouse regulatory T cells (Tregs) from a preparation of naïve CD4+ T cells
Catalog # Availability Size / Price Qty
CDK007
CellXVivo Mouse Treg Cell Differentiation Kit
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Product Details
Procedure
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Reviews (3)

CellXVivo Mouse Treg Cell Differentiation Kit Summary

Kit Summary

For the differentiation of regulatory T Cells (Tregs) from a preparation of naïve CD4+ T cells.

Key Benefits

  • Contains high quality bioactive proteins
  • Yields a highly enriched population of FoxP3+CD25+ Treg cells
  • Provides optimized reagents for inducing Tregs from CD4+ T cells
  • Involves validated and straightforward procedures
  • Does not require specialized instrumentation

 

Why Differentiate Treg Cells In Vitro?

Forkhead Box P3 (FoxP3)+ regulatory T (Treg) cells are a suppressive subset of CD4+ T cells that function to antagonize immune responses. Treg cells have the capacity to prevent potentially damaging autoimmune and protective immune responses, so the number of Treg cells is a crucial determinant of the regulatory burden on the immune system. In vitro differentiation of Treg cells from the larger naïve CD4+ T cell population provides increased numbers of Treg cells to facilitate downstream research.

 

Kit Contents

This kit contains the following optimized proteins and reagents to drive efficient differentiation of naïve CD4+ T cells into FoxP3+CD25+ Treg cells.

  • 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)

 

Stability and Storage

Store the unopened kit at -20 °C. After opening the kit, the Human Anti-Mouse CD3 Antibody, Mouse Treg Reagent 1, and Mouse Treg Reagent 2 may be stored at 2-8 °C under sterile conditions for up to 30 days or at -20 °C to -70 °C in a manual defrost freezer for up to 3 months. Mouse Treg Reagent 3 may be stored at -20 °C to -70 °C in a manual defrost freezer for up to 3 months. Reconstitution Buffer 1, Reconstitution Buffer 2, and 20X Wash Buffer may be stored under sterile conditions for up to 3 months at 2 to 8 °C. Storage conditions are valid if used within the expiration date of the kit.

 

Data Examples
Differentiation of CD4+ T Cells into Treg Cells is Confirmed by Flow Cytometry Analysis of FoxP3 and CD25 Expression
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Differentiation of CD4+ T Cells into Treg Cells Confirmed by FoxP3 and CD25 Expression. Naïve CD4+ T cells were incubated without (A, B) or with (C, D) reagents included in the CellXVivo™ Mouse Treg Cell Differentiation Kit for 5 days. Cells were fixed, permeabilized, and stained using the FlowX Mouse Regulatory T Cell Kit (Catalog # FMC022). Cells were imaged using flow cytometry. Quadrants were set based on isotope-stained samples.

Specifications

Shipping Conditions
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Species
Mouse

Product Datasheets

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

FAQs

No product specific FAQs exist for this product, however you may

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Reviews for CellXVivo Mouse Treg Cell Differentiation Kit

Average Rating: 5 (Based on 3 Reviews)

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