Product Information

Contents: Alexa Fluor® 647 anti-mouse IL-17F (Interleukin-17F)
Catalog Number: 51-7471
Sizes: 25 ug, 100 ug
Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN₃, BSA and/or gelatin for protein stabilization
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: eBio18F10
Isotype: Rat IgG2a

In vitro-differentiated TH17 cells were reactivated with PMA and Ionomycin for 5 hours in the presence of monensin, then stained for CD4 followed by intracellular staining with Alexa Fluor 647 anti-mouse IL-17F clone eBio18F10 (right panel) or isotype control (left panel).

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
12-7471	Phycoerythrin (PE) anti-mouse IL-17F (Interleukin-17F)	488	575	IC Flow
51-7471	Alexa Fluor® 647 anti-mouse IL-17F (Interleukin-17F)	633	668	IC Flow
53-7471	Alexa Fluor® 488 anti-mouse IL-17F (Interleukin-17F)	488	519	IC Flow
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.

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Description

The eBio18F10 antibody reacts with mouse IL-17F. IL-17F is a 37kD 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 (100fold 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

The 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. The antibody can be used at less than or equal to 0.25 µg per million cells in a 100 µl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance as there may be some variation in brightness from one cell type/line to another.

References

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)

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.

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.

Interleukin (IL)-22 and IL-17 are coexpressed by Th17 cells and cooperatively enhance expression of antimicrobial peptides. Liang SC, Tan XY, Luxenberg DP, Karim R, Dunussi-Joannopoulos K, Collins M, Fouser LA. J Exp Med. 2006 Oct 2;203(10):2271-9.

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