Product Pathways - Growth Factors/Cytokines
Human Interleukin-10 (hIL-10) #8903
PhosphoSitePlus® protein, site, and accession data: IL10
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Source
Recombinant human IL-10 (hIL-10) Ser19 - Asn178 (Accession # NM_000572) was produced in E. coli at Cell Signaling Technology.
Molecular Characterization
Recombinant hIL-10 does not have a Met on the amino terminus and has a calculated MW of 18,647. DTT-reduced protein migrates as an 18 kDa polypeptide and non-reduced protein migrates as a 16 kDa polypeptide due to intramolecular cystines. The expected amino-terminal SPGQG of recombinant hIL-10 was verified by amino acid sequencing.
Purity
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hIL-10. All lots are greater than 98% pure.
Bioactivity
The bioactivity of recombinant hIL-10 was determined in a MC/9 cell proliferation assay. The ED50 of each lot is between 0.3-1.3 ng/ml.
Coomassie Gel
The purity of recombinant hIL-10 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIL-10 and staining overnight with Coomassie Blue.
Bioactivity
The proliferation of MC/9 cells treated with increasing concentrations of hIL-10 in the presence of 10 pg/ml mouse IL-4 was assessed. After 72 hours treatment with hIL-10, cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.
Western Blotting
Western blot analysis of extracts from MC/9 cells, untreated or treated with hIL-10 for 20 minutes, using Phospho-Stat3 (Tyr705) (D3A7) Rabbit mAb #9145 (upper) or Stat3 Antibody #9132 (lower).
Endotoxin
Less than 0.01 ng endotoxin/1μg hIL-10.
Formulation
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hIL-10. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Background
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 Stat-3 (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 SOCS-1 (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).
- Pestka, S. et al. (2004) Immunol Rev 202, 8-32.
- Akuffo, H. et al. (1999) Clin Exp Immunol 117, 529-34.
- O'Shea, J.J. and Murray, P.J. (2008) Immunity 28, 477-87.
- Akdis, C.A. and Blaser, K. (2001) Immunology 103, 131-6.
- Akdis, C.A. et al. (2000) FASEB J 14, 1666-8.
- Von Stebut, E. (2000) Eur J Dermatol 17, 115-22.
Application References
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Companion Products
- 9145 Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb
- 9872 Phosphate Buffered Saline (PBS-1X) pH7.2 (Sterile)
For Research Use Only. Not For Use In Diagnostic Procedures.
