Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Mouse His6Interleukin-27 (mHis6IL-27) #5725

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Source

Recombinant mouse His6IL-27 (mHis6IL-27) is a heterodimer that is composed of EBI3 Tyr19-Pro228 (Accession# NP_056581) linked to p28 Phe29-Ser234 (Accession# NP_663611) via the linker GGGSGGGSGGGSGGGS. mHis6IL-27 was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant N-terminally His6-tagged mIL-27 has a calculated MW of 50,551. DTT-reduced and non-reduced protein migrate as 55 kDa polypeptides. The expected amino terminus of recombinant mHis6IL-27 was verified by amino acid sequencing.

Purity

>97% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mHis6IL-27. All lots are greater than 97% pure.

Bioactivity

The bioactivity of mHis6IL-27 was determined in a virus protection assay. The ED50 of each lot is between 1-6 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant mHis6IL-27 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mHis6IL-27 and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The bioactivity of recombinant mHis6IL-27 was determined by induction of MxA mRNA. Hep G2 cells were pre-treated with increasing concentrations of mHis6IL-27 for 5 hr. Induction of MxA gene expression was determined by qPCR.

Bioactivity

Bioactivity

The bioactivity of recombinant mHis6IL-27 was determined in a virus protection assay. Hep G2 cells were pre-treated with increasing concentrations of mHis6IL-27 for 24 hr. Cells were then innoculated with encephalomyocarditis virus (EMCV) and incubated for an additional 24 hr. Surviving cells were fixed and stained with crystal violet and the OD595 was determined.


Western Blotting

Western Blotting

Western blot analysis of extracts from Hep G2 cells, untreated or treated with mHis6IL-27 for 15 min, using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 (upper) or Stat3 Antibody #9132 (lower).

Endotoxin

Less than 0.01 ng endotoxin/1μg mHis6IL-27.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mHis6IL-27. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.

Background

IL-27 is a heterodimer that consists of p28 and EBI3 (1). Antigen presenting cells including monocytes, macrophages, and dendritic cells are the primary sources of IL-27 (1). IL-27 is a pleiotropic cytokine with both pro- and anti-inflammatory activities. IL-27 suppresses TH17 responses and promotes type I regulatory cell differentiation (2-4). In contrast, IL-27 has been shown to play a pro-inflammatory role in a mouse model of T cell-induced colitis (5). IL-27 may also have anti-viral activities (6). IL-27 exerts its effects through a heterodimeric receptor consisting of WSX-1 and gp130 (1,7). IL-27-induced signaling results in Stat1 and Stat3 phosphorylation (1,7).

  1. Pflanz, S. et al. (2002) Immunity 16, 779-90.
  2. Villarino, A.V. et al. (2010) J Immunol 185, 6461-71.
  3. Apetoh, L. et al. (2010) Nat Immunol 11, 854-61.
  4. Murugaiyan, G. et al. (2010) Proc Natl Acad Sci U S A 107, 11495-500.
  5. Cox, J.H. et al. (2011) J Exp Med 208, 115-23.
  6. Bender, H. et al. (2009) Hepatology 50, 585-91.
  7. Pflanz, S. et al. (2004) J Immunol 172, 2225-31.

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