Intracellular staining of human peripheral blood cells stimulated with PMA/Ionomycin in the presence of monensin for 5 hours with Anti-Human CD4 PE (cat. 12-0049) and Mouse IgG1 κ Isotype Control Alexa Fluor® 647 (left) or Anti-Human IL-17A Alexa Fluor® 647 (right). Cells in the lymphocytes gate were used for analysis.
Contents: Anti-Human IL-17A Alexa Fluor® 647 Catalog Number: 51-7178 Concentration: 20 μl (0.125 μg)/test Formulation: Phosphate buffer pH 7.2, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material. Clone: eBio64CAP17 Host/Isotype: Mouse IgG1, κ
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51-7178-71
25 tests
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*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 Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
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.
The eBio64CAP17 has been shown to react to rhesus and marmoset primates.
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 test. A test is defined as the amount (μg)/test 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. 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.
References
Kap YS, van Meurs M, van Driel N, Koopman G, Melief MJ, Brok HP, Laman JD, 't Hart BA. A monoclonal antibody selection for immunohistochemical examination of lymphoid tissues from non-human primates. J Histochem Cytochem. 2009 Dec;57(12):1159-67. (eBio64CAP17, IHC frozen on rhesus and marmoset primates, PubMed)
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.
