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Human IL-17A, amino acids Ile20-Ala155 (Accession # NM_002190) was expressed in E. coli.
The 137 amino acid N-terminal methionylated recombinant protein has a predicted molecular mass of 15,666 Da. This protein exists as a disulfide-linked homodimer. The DTT-reduced protein migrates at approximately 16kDa by SDS-PAGE. The non-reduced protein migrates as a homodimer, at approximately 28kDa by SDS-PAGE.
Purity is >98%, as determined by Coomassie stained SDS-PAGE.
0.22 µm filtered protein solution is in 10mM NaH2PO4, 300mM NaCl, pH 7.2.
Endotoxin level is <0.1 EU/µg (<0.01ng/µg) protein as determined by the LAL method.
10-100µg sizes are bottled at 200µg/ml. 500µg and larger sizes are bottled at the concentration indicated on the vial.
Storage & Handling:
Unopened vial can be stored at -20°C for six months or at -70°C for one year. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 10µg/mL in buffer containing carrier protein such as 1% BSA or HSA or 10% FBS. For long term storage, aliquot into polypropylene vials and store in a manual defrost freezer. Avoid repeated freeze/thaw cycles.
The ED50 is 2 - 4 ng/ml, corresponding to a specific activity 5 - 2.5 x 105 units/mg, as determined by a dose dependent stimulation of normal human dermal fibroblasts production of IL-6.
Induction of IL-6 in human dermal fibroblast by IL-17A.
IL-17A was initially identified from a subtracted cDNA library between closely related murine lymphoid cells and called CTLA-8, and share 58% homology with an open reading frame of the T-lymphotropic Herpesvirus Samirii virus (viral IL-17) (5). IL-17A belongs to a family of cytokines, which has five members; designated IL-17A-F. IL-17 is expressed by a unique lineage of CD4 T cells (Th17) that develop in response to IL-23, in particular under conditions in which Th1 and Th2 development are suppressed. IL-17A shares the greatest homology (55%) with IL-17F. Both IL-17A and IL-17F are produced by Th17 cells. IL-17A and IL-17F can either exist as IL-17A homodimers and IL-17F homodimers or as IL-17A-IL-17F heterodimers (6). IL-17 is a key mediator of autoimmune disorders, including rheumatoid arthritis, psoriasis, inflammatory bowel disease, and asthma, and plays a role in host defense (7).
IL-17A is largely produced by activated memory T lymphocytes, CD4+ T helper cells (Th17), neutrophils, CD8(+), NK, and gamma-delta T cells (3).
IL-17A is a potent regulator of granulopoiesis and neutrophil recruitment under normal and inflammatory conditions. Organ overexpression of IL-17A increases circulating neutrophil numbers and recruitment into the organs by induction of CXCL2, IL-1β, and G-CSF. Il-17A induces CXCL1, CXCL2, CXCL5, and CXCL8 in human epithelial cells. IL-17A also cooperates with TLR ligands, IL-1 beta, and TNF alpha to enhance inflammatory reactions and stimulate production of beta-defensins and other antimicrobial peptides (1, 2).
IL-17A signals through a heteromeric receptor composed of IL-17RA and IL-17RC.
IL-17AR is expressed in epithelial cells, fibroblasts, B and T lymphocytes, myelomonocytic cells, marrow stromal cells, synovial endothelial cells and chondrocytes from arthritic patients express IL-17R (4).
1. Yu J, et al.Front Biosci 13:170-177 2008. 2. Toy D, et al.J. Immunol. 177:36-39 2006. 3. Benghiat FS, et al.Transplant Rev 23:11-18 2009. 4. Honorati MC, et al.Rheumatology 40:522-527 2001. 5. Rouvier E, et al.J. Immunol. 150:5445-5456 1993. 6. Liang SC, et al.J. Immunol. 179:7791-7799 2007. 7. Ouyang W, et al. Immunity 28:454-467 2008.
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