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.
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.
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.
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).
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.
Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
>97% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mHis6IL-27. All lots are greater than 97% pure.
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.
The bioactivity of mHis6IL-27 was determined in a virus protection assay. The ED50 of each lot is between 1-6 ng/ml.
Less than 0.01 ng endotoxin/1μg mHis6IL-27.
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.
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).
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