The production of IL-10 by COLO 205 cells cultured with increasing concentrations of mIL-22 was assessed. Media from cells incubated with mIL-22 for 48 hours was collected and assayed for IL-10 by ELISA and the OD450-OD650 was determined.
The purity of recombinant mIL-22 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIL-22 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from HT-29 cells, untreated or treated with mIL-22 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 mIL-22. 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 mIL-22. All lots are greater than 98% pure.
Recombinant mIL-22 contains no "tags" and the nonglycosylated protein has a calculated MW of 16,640. 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 LPVNT of recombinant mIL-22 was verified by amino acid sequencing.
The bioactivity of recombinant mIL-22 was determined by its ability to induce IL-10 production by COLO 205 cells. The ED50 of each lot is between 0.2-0.8 ng/ml.
Less than 0.01 ng endotoxin/1μg mIL-22.
Recombinant mouse IL-22 (mIL-22) Leu34-Val179 (Accession #NP_058667) was expressed in human 293 cells at Cell Signaling Technology.
IL-22, a member of the IL-10 family (1,2), is expressed by Th17 CD4+ T cells, activated T cells, Th1 cells and NK cells (3). Expression of IL-22 in combination with a select group of cytokines defines a subset of Th cells(4). IL-22 induces proinflammatory responses, drives production of antimicrobial peptides, and is involved in tissue-repair and wound-healing responses (1). The IL-22 receptor is a heterodimer of IL-22R1 and IL-10R2 (5). Expression of IL-22R is restricted to tissue-resident cells, particularly those of epithelial origin, whereas the IL-10R2 chain is expressed in many more cell types. IL-22 induces phosphorylation of Jak1 and Tyk2, leading to activation of Stat3 and, to a lesser extent, Stat1 and Stat5 (1). IL-22 responses can involve activation of the MEK-ERK-RSK, JNK-SAPK, and p38 pathways (1). Elevated levels of IL-22 have been associated with Crohn's disease and rheumatoid arthritis. IL-22 plays an essential role in host response to the pulmonary pathogen Klebsiella pneumoniae (6).
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