CD4-enriched human peripheral blood cells were polarized under Th17 conditions (with Human IL-23 Recombinant Protein (Product # 14-8239)) for 10 days. Cells were restimulated with Protein Transport Inhibitor Cocktail (Product # 00-4980) (left) or Cell Stimulation Cocktail plus Protein Transport Inhibitors (Product # 00-4975) (right) for 6 hours. Cells were fixed and permeabilized using the FIntracellular Fixation and Permeabilization Buffer Set (Product # 88-8824) and then intracellularly stained with Anti-Human CD4 APC (Product # 17-0049) and Anti-Human IL-17AF eFluor® 450. Viable cells, as determined by Fixable Viability Dye eFluor® 780 (Product # 65-0865), were used for analysis.
|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin, 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||5 µL (0.5 µg)/test|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Miscellaneous PubMed (MISC)||See 3 publications below|
Description: The 20LJS09 monoclonal antibody reacts with human interleukin (IL)-17AF heterodimer. This antibody specifically reacts with this heterodimer and has no reactivity for IL-17A or IL-17F homodimers. IL-17A and IL-17F are well-characterized homodimeric cytokines secreted by T helper 17 (Th17) cells, gamma-delta T cells and several subsets of innate lymphoid cells. Somewhat less appreciated, IL-17A and IL-17F subunits can also form the heterodimer, IL-17AF. Together, these three dimers signal through the IL-17RA/IL-17RC receptor complex to mediate immune responses at mucosal interfaces and are found at lesion sites in inflammatory bowel disease, asthma, atopic dermatitis and rheumatoid arthritis.
Applications Reported: This 20LJS09 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This 20LJS09 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of in vitro-polarized human Th17 cells. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) 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 10^5 to 10^8 cells/test.
eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome.
Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser
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 receptor. IL-17A binds to a receptor that binds also to HVS13 (viral IL-17) and to CTLA-8.This recombinant protein is E. coli expressed. Insect cell expressed Human IL-17A Recombinant Protein (cat. 14-8180) is also available.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||Identification of an interleukin 17F/17A heterodimer in activated human CD4+ T cells.||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||The Journal of biological chemistry (282:13447)||2007|
|Not Applicable||Not Cited||A novel heterodimeric cytokine consisting of IL-17 and IL-17F regulates inflammatory responses.||Chang SH,Dong C||Cell research (17:435)||2007|
|Not Applicable||Not Cited||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||The Journal of experimental medicine (203:2271)||2006|