T helper type 2 (Th2) cells are a lineage of CD4+ effector T cells that provide host protection against intestinal parasites and extracellular bacteria. In addition, they provide support for B cell-dependent humoral responses. Pathological Th2 cell activity is a hallmark of allergic inflammation and asthma. Differentiation of CD4+ effector cells into the Th2 lineage is promoted by cytokines such as IL-4 in combination with either IL-2, IL-7, or TSLP. Th2 cells secrete IL-4, IL-5, IL-9, IL-13, and IL-17E/IL-25. In vitro differentiation of mouse Th2 cells from the larger naïve CD4+ T cell population provides increased numbers of Th2 cells to facilitate downstream research. The CellXVivo™ Mouse Th2 Cell Differentiation Kit contains all necessary components to differentiate mouse naïve CD4+ T cells into Th2 polarized cells.
This kit contains the following reagents for the ex vivo differentiation of mouse Th2 cells.
Hamster Anti-Mouse CD3, Th2
Rat Anti-Mouse CD28, Th2
Mouse Th2 Reagent 1
Mouse Th2 Reagent 2
Mouse Th2 Reagent 3
Reconstitution Buffer 1
Reconstitution Buffer 2
20X Wash Buffer
The quantity of the components in the kit is sufficient to differentiate approximately 4 x 106 naïve CD4+ T cells, and generate 80 x 106 T cells, of which >40% are IL-4+ Th2 polarized cells.
Note: Results may vary due to strain, age, and/or the health of the mice used for isolation.
Stability and Storage
Store the unopened kit at ≤-20 °C. Do not use past the kit expiration date.
FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
This reagent should not be used beyond the expiration date indicated on the label.
Results may vary due to variations among cells derived from different donors.
Intracellular Cytokine Staining of Differentiated Mouse Th2 Cells. Flow cytometry data of mouse naïve CD4+ T cells (A, C) and differentiated Th2 cells (B, D) generated using reagents included in this kit. After 6 days of differentiation, naïve CD4+ cells or differentiated Th2 cells were stimulated with Cell Activation Cocktail (Tocris®, Catalog # 5476) and stained with conjugated Anti-Mouse IL-4 (Clone 11B11), Anti-Mouse IFN-gamma (Catalog # IC485P), and Anti-Mouse IL-17A (Novus Biologicals®, Catalog # NBP1-72027) Monoclonal Antibodies. Quadrants were set based on isotype-stained controls.
Th2-differentiated Mouse CD4+ Cells Secrete High Levels of IL-4. Mouse naïve CD4+ T cells were differentiated for 6 days using the reagents included in this kit. On day 6 of differentiation cells were harvested and stimulated with Anti-Mouse CD3 and Anti-Mouse CD28 overnight. Cell culture supernatant was collected and cytokine secretion determined using the Mouse IL-4 Quantikine® ELISA Kit (Catalog # M4000B), the Mouse IFN-gamma Quantikine® ELISA Kit (Catalog # MIF00), and the Mouse IL-17 Quantikine® ELISA Kit (Catalog # M1700).
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