Intracellular staining of day 9 Th17-polarized Balb/c splenocytes with eFluor® 450 anti-mouse CD4 (RM4-5) (Pacific Blue® replacement) (cat. 48-0042) and 0.125 μg of APC anti-mouse IL-21. Cells were polarized for 9 days and then treated for 5 hours with Brefeldin A alone (cat. 00-4506) (left) or PMA, ionomycin and Brefeldin A (right). Cells in the lymphocyte gate were used for analysis.
Contents: Allophycocyanin (APC) anti-mouse IL-21 (Interleukin-21, IL21) Catalog Number: 17-7211 Concentration: 0.2 mg/mL Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary Storage Conditions: Store at 2-8°C. DO NOT FREEZE. LIGHT-SENSITIVE MATERIAL. Clone: FFA21 Host/Isotype: Rat IgG2a, k
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17-7211-80
25 ug
17-7211-82
100 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.
Description
This FFA21 monoclonal antibody reacts with mouse interleukin-21 (IL-21). IL-21 is a 17 kDa immunomodulatory cytokine produced mainly by NKT, T helper (Th) 17 and T follicular helper (TFH) cells. In TFH cells, IL-21 expression leads to autocrine signaling through the IL-21 receptor (IL-21R) and STAT3, which leads to additional transcriptional activation by Bcl6. As with IFNγ for Th1, IL-4 for Th2 and IL-17A for Th17, IL-21 is critical for TFH cell effector function. This cytokine plays a role in T cell-dependent B cell differentiation into plasma cells and memory cells, stimulation of IgG production and induction of apoptotic signaling in naļve B cells.
In Th17 cells, IL-21 expression and autocrine feedback through STAT3, IRF4 and RORγt lead to upregulation of the IL-23R, thereby preparing Th17 cells for maturation and maintenance by the inflammatory cytokine IL-23. While upregulating IRF4 and RORγt, IL-21 also mediates the downregulation of Foxp3. High levels of IL-21 are present in chemically-induced colitis models. IL-21-deficient mice are protected from developing colitis upon chemical treatment by their inability to upregulate Th17-associated molecules.
Preliminary data suggest that clone FFA21 recognizes a different epitope than anti-mouse IL-21 clone mhalx21 (cat. 51-7213).
Applications Reported
For research use only, not for diagnostic or therapeutic use. This FFA21 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested
This FFA21 antibody has been tested on Th17-polarized splenocytes restimulated with PMA, Ionomycin and Brefeldin A for 5 hours. This can be used at less than or equal to 0.125 μg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µl. Cell number should be determined empirically but can range from 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Konforte D, Paige CJ. Interleukin-21 regulates expression of the immediate-early lytic cycle genes and proteins in Epstein-Barr Virus infected B cells. Virus Res. 2009 Sep;144(1-2):339-43.
Pot C, Jin H, Awasthi A, Liu SM, Lai CY, Madan R, Sharpe AH, Karp CL, Miaw SC, Ho IC, Kuchroo VK. Cutting edge: IL-27 induces the transcription factor c-Maf, cytokine IL-21, and the costimulatory receptor ICOS that coordinately act together to promote differentiation of IL-10-producing Tr1 cells. J Immunol. 2009 Jul 15;183(2):797-801.
Elsaesser H, Sauer K, Brooks DG. IL-21 is required to control chronic viral infection. Science. 2009 Jun 19;324(5934):1569-72.
Bauquet AT, Jin H, Paterson AM, Mitsdoerffer M, Ho IC, Sharpe AH, Kuchroo VK. The costimulatory molecule ICOS regulates the expression of c-Maf and IL-21 in the development of follicular T helper cells and TH-17 cells. Nat Immunol. 2009 Feb;10(2):167-75.
Nurieva R, Yang XO, Martinez G, Zhang Y, Panopoulos AD, Ma L, Schluns K, Tian Q, Watowich SS, Jetten AM, Dong C. Essential autocrine regulation by IL-21 in the generation of inflammatory T cells. Nature. 2007 Jul 26;448(7152):480-3.
