Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Interleukin-17A (hIL-17A) #8928

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Source

Recombinant human IL-17A (hIL-17A) Ile20-Ala155 (Accession #NP_002181) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant hIL-17A contains no "tags" and the nonglycosylated protein has a calculated MW of 15,535. DTT-reduced protein migrates as a 16-24 kDa polypeptide. Heterogeneity in SDS PAGE is due to glycosylation. The non-reduced cystine-linked homodimer migrates as a 28-37 kDa protein. The expected amino-terminal IVKAG of recombinant hIL-17A was verified by amino acid sequencing.

Purity

>97% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hIL-17A. All lots are greater than 97% pure

Bioactivity

The bioactivity of recombinant hIL-17A was determined by its ability to induce IL-6 production by primary human fibroblasts. The ED50 of each lot is between 1.5 - 3.5 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hIL-17A was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIL-17A and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The production of IL-6 by primary human fibroblasts cultured with increasing concentrations of human IL-17A was assessed. Media from cells incubated with IL-17A for 48 hours was collected and assayed for IL-6 by ELISA and the OD450 - OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from human foreskin fibroblasts untreated or treated with hIL-17A for 15 minutes, using Phospho-p38 MAPK (Thr180/Tyr182) (3D7) Rabbit mAb #9215 (upper) and p38 MAPK Antibody #9212 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hIL-17A.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hIL-17A. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.

Background

IL-17A is a cystine-linked homodimeric pro-inflammatory cytokine produced by Th17 cells, a distinct CD4+ T cell lineage (1,2). IL-17A stimulates the production of the pro-inflammatory cytokines IL-1β, TNF-α, and IL-6. IL-17A also induces production of the neutrophil chemoattractants IL-8, CXCL1, and CXCL6 thereby bridging adaptive and innate immunity (1,2). IL-17A is intimately involved in mucosal immunity against bacterial infections (1,3) and has a putative role in some autoimmune disorders (1,4). IL-17A effects appear to be exerted primarily through binding to the IL-17RA (5). IL-17A binding induces production of cytokines, chemokines and other proteins through activation of the Erk1/2 MAP kinase, PI3K/Akt, p38, and NF-κB pathways (3,4, 6). Phosphorylation of some Jaks and Stats has been observed.

  1. Kolls, J.K. and Lindén, A. (2004) Immunity 21, 467-76.
  2. Liang, S.C. et al. (2006) J Exp Med 203, 2271-9.
  3. Dubin, P.J. and Kolls, J.K. (2008) Immunol Rev 226, 160-71.
  4. Zrioual, S. et al. (2009) J Immunol 182, 3112-20.
  5. Wright, J.F. et al. (2008) J Immunol 181, 2799-805.
  6. Rahman, M.S. et al. (2006) J Immunol 177, 4064-71.

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