The bioactivity of recombinant hIL-29 was determined in a virus protection assay. Hep G2 cells were pre-treated with increasing concentrations of hIL-29 for 24 hours. Cells were then innoculated with encephalomyocarditis virus (EMCV) and incubated for an additional 24 hours. Surviving cells were then fixed and stained with crystal violet and the OD595 was determined.
The purity of recombinant hIL-29 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIL-29 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from RPMI 8226 cells untreated or treated with hIL-29 for 15 minutes, 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 hIL-29. 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.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hIL-29. All lots are greater than 98% pure.
Recombinant hIL-29 contains no "tags" and the nonglycosylated protein has a calculated MW of 20,018. DTT-reduced and non-reduced protein migrate as 31 kDa polypeptides. Lower mobility and heterogeneity in SDS-PAGE are due to glycosylation. The expected amino-terminal GPVPT of recombinant hIL-29 was verified by amino acid sequencing.
The bioactivity of hIL-29 was determined in a virus protection assay. The ED50 of each lot is between 0.40-1.45 ng/ml.
Less than 0.01 ng endotoxin/1μg hIL-29.
Recombinant human IL-29 (hIL-29) Gly20-Thr200 (Accession #NP_742152) was expressed in human 293 cells at Cell Signaling Technology.
IL-29 is a member of the IFNλ family of cytokines and shares many functions with the type I family of interferons (1,2). IL-29 is produced by monocytes and dendritic cells in response to viral infection and stimulation with toll-like receptor ligands (1-3). IL-29 is a potent antiviral cytokine. The antiviral activities of IL-29 include the upregulation of MHC Class I expression on the cell surface, and the expression of PKR, MxA, and 2’, 5'-OAS (1,2). The IL-29 receptor consists of IL-28Rα and IL-10Rβ is expressed on most non-hematopoietic cells (3). IL-29 binding induces the phosphorylation of Stats 1, 2 and 3 (1,3).
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