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

Contents: Alexa Fluor® 647 anti-human IL-17A (Interleukin-17A)
Catalog Number: 51-7178
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: eBio64CAP17
Isotype: Mouse IgG1, κ

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51-7178-71

25 tests

Human PBMCs were stimulated with TPA/ionomycin in the presence of monensin for 5 hours. The cells were fixed and permeabilized and intracellularly stained with PE anti-human CD4, clone RPA-T4, (cat. 12-0049) and Alexa 647 anti-human IL-17A. Isotype control on the left and anti-IL-17A on the right. Cells in the lymphocytes gate were used for analysis.

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. The eBio64CAP17 antibody has been reported for use as the capture antibody in human IL-17A ELISA and ELISPOT assays, for neutralization of IL-17A bioactivity, and for intracellular staining and flow cytometric analysis of IL-17A-producing cells from activated cell cultures.

Applications Tested

The Alexa Fluor® 647-conjugated eBio64CAP17 antibody has been pre-titrated and tested for intracellular immunofluorescent staining and flow cytometry. This can be used at 20 µl (0.125 µg) per million cells in a 100 µl total staining volume. IL-17A-expressing cells were identified within the CD4 population of human PBMCs which were stimulated with TPA/ionomycin for 5 hrs in the presence of monensin.

Related Products

Cat. 51-4714  Alexa Fluor® 647 Mouse IgG1, K Isotype Control
Cat. 88-7176  Human IL-17A (Interleukin-17A, IL17A) ELISA Ready-SET-Go!
Cat. 13-7179  Biotin anti-human IL-17A (Interleukin-17A, IL17A) (clone eBio64DEC17)
Cat. 88-7876  Human IL-17A (Interleukin-17A, IL17A) ELISPOT Ready-Set-Go!
Cat. 88-7976  Human IL-17A (Interleukin-17A, IL17A) ELISA Ready-SET-Go! Kit (with Pre-Coated Plates)

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.