Alexa Fluor® 647 anti-mouse IL-17A antibody: Alexa Fluor® 647 conjugate Interleukin-17A

Contents: Alexa Fluor® 647 anti-mouse IL-17A (Interleukin-17A)
Catalog Number: 51-7172
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃, 0.1% gelatin
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: eBioTC11-18H10.1
Isotype: Rat IgG1, κ

Prices for This Product*
Cat. No.	Size	Price	Qty	Action
51-7172-80	25 ug	$195

*International customers: Please contact your distributor for region specific pricing.

Intracellular staining of mouse IL-17A. CD4+CD25- T cells purified from C57BL/6 mouse splenocytes were cultured with (day 10) bone marrow-derived dendritic cells in the presence of anti-CD3, anti-IL-4, anti-IFN-γ, recombinant TGF-β, and recombinant IL-6 for 4 days. Cells were then reactivated with PMA/Ionomycin in the presence of monensin for 5 hrs. Cells were harvested, fixed, permeabilized, and stained with 0.25 ug of Alexa Fluor® 647 anti-mouse IL-17A and 0.5 ug of PE anti-mouse IFN-γ.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
51-7172	Alexa Fluor® 647 anti-mouse IL-17A (Interleukin-17A, IL17A)	633	668	IC Flow
53-7172	Alexa Fluor® 488 anti-mouse IL-17A (Interleukin-17A, IL17A)	488	519	IC Flow

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The eBioTC11-18H10.1 antibody reacts with mouse IL-17A. The eBioTC11-18H10.1 antibody is a neutralizing antibody. 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-γ and IL-4, have each been found to negatively regulate the generation of these Th-17 cells.

Applications Reported

For research use only, not for diagnostic or therapeutic use. The eBioTC11-18H10.1 antibody has been reported for use as the capture antibody in mouse IL-17A ELISA and ELISPOT assay, for neutralization of IL-17A bioactivity, and for intracellular staining of IL-17A-producing cells.

Applications Tested

The fluorochrome-conjugated eBioTC11-18H10.1 antibody has been tested by intracellular staining for flow cytometric analysis and can be used at less than or equal to 0.50 µg/million cells/100 ul staining volume.

It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. The Functional Grade Purified eBioTC11-18H10.1 antibody has been tested by LAL assay to verify low endotoxin and has been tested for neutralization of IL-17A bioactivity and for ELISPOT capture.

The Affinity Purified eBioTC11-18H10.1 antibody has been tested as the capture antibody in a sandwich ELISA for measurement of mouse IL-17A protein levels, in combination with the biotinylated eBioTC11-8H4 antibody (13-7171) for detection and recombinant mouse IL-17A (14-8171) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 0.5 – 2.0 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 2000 pg/ml - 15 pg/ml should be included in each ELISA plate.

References

Mangen PR, Harrington LE, O'Quinn DB, Helms WS, Bullard DC, Elson CO, Hatton RD, Wahl SM, Schoeb TR, Weaver CT. 2006. Transforming growth factor-beta induces development of the Th-17 lineage. Nature. Letters. Advance Online Publication. 30 April, 2006. [Intracellular staining]
Harrington LE, Hatton RD, Mangan PR, Turner H, Murphy TL, Murphy KM, Weaver CT. 2005. Interleukin 17–producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nature Immunol 6: 1123-1132. [ELISA, Intracellular Staining]
Park H, Li X, Yang XO, Chang SH, Nurieva R, Wang Y-H, Wang Y, Hood L, Zhu S, Tian Q, Dong C. 2005. A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nature Immunol 6: 1133-1141. [ELISA, Intracellular Staining]
Nekrasova T, Shive C, Gao Y, Kawamura K, Guardia R, Landreth G, Forsthuber TG. 2005. ERK1-deficient mice show normal T cell effector function and are highly susceptible to experimental autoimmune encephalomyelitis. J Immunol. 175: 2374-80. [ELISPOT]
Schubert D, Maier B, Morawietz L, Krenn V, Kamradt T. 2004. Immunization with Glucose-6-Phosphate Isomerase Induces T Cell-Dependent Peripheral Polyarthritis in Genetically Unaltered Mice. J Immunol 172: 4503-4509. [Intracellular Staining]
Infante-Duarte C, Horton HF, Byrne MC, Kamradt T. 2000. Microbial Lipopeptides Induce the Production of IL-17 in Th Cells. J Immunol. 165: 6107-6115. [Intracellular Staining]
Hofstetter HH, Ibraihim SM, Koczan D, Kruse N, Weishaupt A, Toyka KV, Gold R. 2005. Therapeutic efficacy of IL-17 neutralization in murine experimental autoimmune encephalomyelitis. Cell Immunol. [Dec 27 2005 Epub ahead of print; ELISPOT]

Related Products

Cat. 13-7171 Biotin anti-mouse IL-17A (Interleukin-17A, IL17A) (clone eBioTC11-8H4)
Cat. 11-7311 FITC anti-mouse IFNg (Interferon-gamma, IFN-g) (clone XMG1.2)

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.