Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Mouse Interleukin-17A (mIL-17A) #5227

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Source

Recombinant mouse IL-17A (mIL-17A) Thr22-Ala158 (Accession #NP_034682) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant mIL-17A contains no "tags" and the nonglycosylated protein has a calculated MW of 15,377. DTT-reduced protein migrates as a 15-22 kDa polypeptide. Heterogeneity in SDS PAGE is due to glycosylation. The non-reduced cystine-linked homodimer migrates as a 28-36 kDa protein. The expected amino-terminal TVKAA of recombinant mIL-17A was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mIL-17A. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant mIL-17A was determined by its ability to induce mouse IL-6 production by 3T3 MEFs WT. The ED50 of each lot is between 0.4-1.4 ng/ml.

Coomassie Gel

Coomassie Gel

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

Bioactivity

Bioactivity

The production of mouse IL-6 by 3T3 MEFs WT cultured with increasing concentrations of mIL-17A was assessed. Media from cells incubated with mIL-17A for 24 hours was collected and assayed for mouse IL-6 by ELISA and the OD450-OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from 3T3 MEFs WT untreated or treated with mIL-17A for 10 minutes, using Phospho-IκB-α (Ser32) (14D4) Rabbit mAb #2859 (upper) and IκB-α (L35A5) Mouse mAb #4814 (lower).


Endotoxin

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

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mIL-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, TNFα, 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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