The proliferation of MC/9 cells treated with increasing concentrations of mIL-10 in the presence of 1 pg/ml mouse IL-4 (#5208) was assessed. After 72 hour treatment with mIL-10 cells, were incubated with a tetrazolium salt and the OD450-OD650 was determined.
The purity of recombinant mIL-10 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIL-10 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from MC/9 cells, untreated or treated with mIL-10 for 20 minutes, using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 (upper) or Stat3 Antibody #9132 (lower).
With carrier: A 0.22 μm filtered solution of 0.33 mg/ml mIL-10 in PBS, pH 7.2 containing 20 μg BSA per 1 μg mIL-10. Carrier free: A 0.22 μm filtered solution of 0.33 mg/ml mIL-10 in PBS, pH 7.2.
Stable at -20°C for 1 year after receipt. Maintain sterility. Do not store frozen.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mIL-10. All lots are greater than 98% pure.
Less than 0.01 ng endotoxin/1 μg mIL-10.
The bioactivity of recombinant mIL-10 was determined in a MC/9 cell proliferation assay. The ED50 of each lot is between 0.3-1.3 ng/ml.
Recombinant mIL-10 does not have a Met on the amino terminus and has a calculated MW of 18,976. DTT-reduced and non-reduced protein migrate as 19 kDa polypeptides. The expected amino-terminal SRGQY of recombinant mIL-10 was verified by amino acid sequencing.
Recombinant mouse IL-10 (mIL-10) Ser19 - Ser178 (Accession # NP_034678) was produced in E. coli at Cell Signaling Technology.
IL-10 is an anti-inflammatory cytokine that is produced by T cells, NK cells, and macrophages (1,2). IL-10 initiates signal transduction by binding to a cell surface receptor complex consisting of IL-10 RI and IL-10 RII (1). Binding of IL-10 leads to the activation of Jak1 and Tyk2, which phosphorylates Stat3 (1,3). The anti-inflammatory activity of IL-10 is due to its ability to block signaling through other cytokine receptors, notably IFNγ receptor, by upregulating expression of SOCS1 (1,3). In addition, IL-10 promotes T cell tolerance by inhibiting tyrosine phosphorylation of CD28 (4,5). IL-10 is an important negative regulator of the immune response, which allows for maintenance of pregnancy (1). In contrast, increased IL-10 levels contribute to persistent Leishmania major infections (6).
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