Product Information

Contents: Affinity Purified anti-human IL-17A (Interleukin-17A)
Catalog Number: 14-7178
Sizes: 50 ug, 500 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
Clone: eBio64CAP17
Isotype: Mouse IgG1, κ

Immunoprecipitation of IL-17A/F heterodimer with eBio64CAP17 followed by immunoblot with H17F10A7 anti-IL-17F antibody. PBMC (1 and 4), in vitro-polarized Th17 (2 and 5), or PMA and Ionomycin-restimulated Th17 (3 and 6) were loaded. Direct lysates (lanes 1-3) are on the left and IP samples (lanes 4-6) are on the right.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-7178	Affinity Purified anti-human IL-17A (Interleukin-17A, IL17A)	N/A	N/A	ELISA cap ELISPOT cap IC Flow IP
16-7178	Functional Grade* Purified anti-human IL-17A (Interleukin-17A, IL17A)	N/A	N/A	ELISA cap ELISPOT cap NU
12-7178	Phycoerythrin (PE) anti-human IL-17A (Interleukin-17A, IL17A)	488	575	IC Flow
51-7178	Alexa Fluor® 647 anti-human IL-17A (Interleukin-17A, IL17A)	633	668	IC Flow
*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.

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Description

The eBio64CAP17 antibody reacts with human IL-17A; the antibody has been reported to cross react with Rhesus monkey IL-17A, as verified by intracellular staining experiments. The eBio64CAP17 antibody is a neutralizing antibody. Reactivity of the eBio64CAP17 antibody with other IL-17 family members has not been evaluated. Interleukin-17A (IL-17A) is a CD4+ T cell-derived cytokine that promotes inflammatory responses in cell lines and is elevated in rheumatoid arthritis, asthma, multiple sclerosis, psoriasis, and transplant rejection. The cDNA encoding human IL-17A was isolated from a library of CD4+ T cells; the encoded protein exhibits 72 percent amino acid identity with HVS13 , an open reading frame from a T lymphotropic Herpesvirus saimiri, and 63 percent with mouse CTLA-8 (cytotoxic T-lymphocyte associated antigen-8). Human IL-17A exists as glycosylated 20-30 kD homodimers. High levels of IL-17A homodimer are produced by activated peripheral blood CD4+ T-cells. IL-17A enhances expression of the intracellular adhesion molecule-1 (ICAM-1) in human fibroblasts. Human IL-17A also stimulates epithelial, endothelial, or fibroblastic cells to secrete IL-6, IL-8, G-CSF, and PGE2. In the presence of human IL-17A, fibroblasts can sustain the proliferation of CD34+ hematopoietic progenitors and induce maturation into neutrophils. Mouse, rat, and human IL-17A can induce IL-6 secretion in mouse stromal cells, indicating that all homologs can recognize the mouse IL-17A receptor.

IL-23-dependent, IL-17A-producing CD4+ T cells (Th-17 cells) have been identified as a unique subset of Th cells that develops along a pathway that is distinct from the Th1- and Th2- cell differentiation pathways. The hallmark effector molecules of Th1 and Th2 cells, e.g., IFN-g and IL-4, have each been found to negatively regulate the generation of these Th-17 cells.

Additionally, activated human CD4+ T cells have been found to produce the IL-17A/F heterodimer, as well as the corresponding homodimers. In comparing the relative potency of IL-17A, IL-17F, and IL-17A/F, all three were found to induce GRO-a secretion; IL-17A was most potent, followed by IL-17A/F heterodimer, then IL-17F (100fold lower than IL-17A). eBio64CAP17 can be used to detect IL-17 heterodimers by immunoprecipitation followed by immunoblot with H17F10A7 anti-IL17F monoclonal antibody.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This eBio64CAP17 antibody has been reported for use in intracellular staining followed by flow cytometric analysis, immunoprecipitation, ELISA, and ELISPOT. (Fluorochrome conjugated eBio64CAP17 is recommended for use in intracellular flow cytometry.)

Applications Tested

The Affinity Purified eBio64CAP17 antibody has been tested as the capture antibody in a sandwich ELISA for measurement of human IL-17A protein levels, in combination with the biotinylated eBio64DEC17 antibody (13-7179) for detection and recombinant human IL-17A (14-8179) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 0.5 – 2.0 ug µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/ml - 8 pg/ml should be included in each ELISA plate.

The Functional Grade Purified eBio64CAP17 antibody has been tested by LAL assay to verify lowest endotoxin levels and has been tested for neutralization of IL-17A bioactivity and for ELISPOT capture.

It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

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