Intracellular staining of EL-4 cells stimulated with PMA/Ionomycin in the presence of monensin with Anti-Mouse IL-17A Alexa Fluor® 647 and 0.25 µg of Rat IgG2a κ Isotype Control Alexa Fluor® 488 (cat. 53-4321) (left) or 0.25 µg of Anti-Mouse IL-17F Alexa Fluor® 488 (right).
Contents: Anti-Mouse IL-17F Alexa Fluor® 488 Catalog Number: 53-7471 Concentration: 0.5 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: eBio18F10 Host/Isotype: Rat IgG2a
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53-7471-80
25 ug
53-7471-82
100 ug
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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 Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The eBio18F10 antibody reacts with mouse IL-17F. IL-17F is a 37 kD homodimer of the IL-17 family and a signature Th17 marker. Of all the six IL-17 family members, IL-17F and IL-17A share the strongest homology (50% amino acid identity), and the two genes are located in the same chromosomal region. Recent studies have demonstrated coordinated regulation of IL-17A and IL-17F during Th17 differentiation. Expression of IL-17F and IL-17A has been detected in activated human peripheral blood lymphocytes, specifically by activated human CD4+ T cells. In addition to IL-17A, differentiated Th17 cells also produce IL-17F and IL-22 upon re-activation. Like IL-17A, IL-17F has been linked with inflammatory diseases. IL-17F and IL-17A expression has been observed in tissue samples from various autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis, psoriasis, inflammatory bowel disease, and asthma. IL-17F treatment of airway epithelium, vein endothelial cells, and fibroblasts has been reported to induce expression of IL-6, IL-8, GRO-α, ENA-78, TGF-β, MCP-1, G-CSF, GM-CSF, and ICAM-1.
Like IL-17A, IL-17F is a disulfide-linked homodimeric glycoprotein. The IL-17F homodimer includes a classical cysteine knot motif, which is found also in the TGF-β, BMP, and NGF superfamilies. The presence of the cysteine knot motif suggested the possibility of a heterodimeric structure, as was reported for TGF-β and inhibin/activin. Recent reports confirm that co-expression of IL-17F and IL-17A in HEK293 cells results in the formation of biologically active IL-17F/IL-17A heterodimers, in addition to the IL-17F homodimers and IL-17A homodimers. Moreover, activated human CD4+ T cells were found to produce the IL-17A/F heterodimer, along with the corresponding homodimers. In comparing the relative potency of IL-17A, IL-17F, and IL-17A/F, all three were found to induce GRO-α secretion; IL-17A was most potent, followed by IL-17A/F heterodimer, then IL-17F (100-fold lower than IL-17A). In the mouse, the IL-17A/F heterodimer (alone or in synergy with TNF-α) was found to regulate the expression of IL-6 and KC (mouse homolog of human GRO-α); this was found to be dependent on IL-17RA and TRAF6.
For intracellular staining and flow cytometric analysis with direct conjugates of anti-mouse IL-17F, use of the Foxp3 buffer system (cat. 00-5523) is recommended. Other buffers may yield varying results. For more information, please contact technical support at tech@ebioscience.com.
Applications Reported
For research use only, not for diagnostic or therapeutic use. The eBio18F10 antibody has been reported useful for intracellular staining and flow cytometric analysis.
Applications Tested
This eBio18F10 antibody has been tested by intracellular staining and flow cytometric analysis of in vitro-differentiated mouse Th17 cell cultures and on the EL-4 cell line. This can be used at less than or equal to 0.5 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
Chang SH, Dong C. A novel heterodimeric cytokine consisting of IL-17 and IL-17F regulates inflammatory responses. Cell Res. 2007 May;17(5):435-40.
Lee YK, Turner H, Maynard CL, Oliver JR, Chen D, Elson CO, Weaver CT. Late developmental plasticity in the T helper 17 lineage. Immunity. 2009 Jan;30(1):92-107. (18F10, IC Flow, Pubmed)
Liang SC, Tan XY, Luxenberg DP, Karim R, Dunussi-Joannopoulos K, Collins M, Fouser LA. Interleukin (IL)-22 and IL-17 are coexpressed by Th17 cells and cooperatively enhance expression of antimicrobial peptides. J Exp Med. 2006 Oct 2;203(10):2271-9.
Wright JF, Guo Y, Quazi A, Luxenberg DP, Bennett F, Ross JF, Qiu Y, Whitters MJ, Tomkinson KN, Dunussi-Joannopoulos K, Carreno BM, Collins M, Wolfman NM. Identification of an interleukin 17F/17A heterodimer in activated human CD4+ T cells. J Biol Chem. 2007 May 4;282(18):13447-55.
Legal
Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.