Recombinant Human IL-10, Animal Free Protein

Animal-Free™ Recombinant Human IL-10 (Catalog # AFL1064) was measured in a cell proliferation assay using MC/9-2 mouse mast cells. The ED50 for this effect is 0.15-0.75 ng/mL.

2 μg/lane of Animal-Free™ Recombinant Human IL-10 Protein (Catalog # AFL1064) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® blue staining, showing bands at 18 kDa and 16 kDa, respectively.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human IL-10, Animal Free Protein Summary

• Additional Information: Intended for preclinical researchers who may transition to GMP IL-10 for their clinical work
• Details of Functionality: Measured in a cell proliferation assay using MC/9‑2 mouse mast cells. Thompson-Snipes, L. et al. (1991) J. Exp. Med. 173:507. The ED₅₀ for this effect is 0.0750-0.750 ng/mL.

  The specific activity of recombinant human IL-10 is >2.50 x 10⁶ units/mg, which is calibrated against the human IL-10 WHO Reference Reagent (NIBSC code: 93/722).

• Source: E. coli-derived human IL-10 protein
  Ser19-Asn178, with an N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
• Accession #: P22301.1
• N-terminal Sequence: Met
• Structure / Form: Noncovalently-linked homodimer
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Gene: IL10
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 19 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 18 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining
• Reconstitution Instructions: Reconstitute at 100-500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human IL-10, Animal Free Protein

• CSIF
• CSIFMGC126450
• Cytokine synthesis inhibitory factor
• GVHDS
• IL10
• IL-10
• IL10A
• IL-10MGC126451
• interleukin 10
• interleukin-10
• TGIF

Background

Interleukin 10, also known as cytokine synthesis inhibitory factor (CSIF), is the charter member of the IL‑10 family of alpha ‑helical cytokines that also includes IL‑19, IL‑20, IL‑22, IL‑24, and IL‑26/AK155 (1, 2). IL‑10 is secreted by many activated hematopoietic cell types as well as hepatic stellate cells, keratinocytes, and placental cytotrophoblasts (2-5). Mature human IL‑10 shares 72%-86% amino acid sequence identity with bovine, canine, equine, feline, mouse, ovine, porcine, and rat IL‑10. Whereas human IL‑10 is active on mouse cells, mouse IL‑10 does not act on human cells (6, 7). IL‑10 is a 178 amino acid molecule that contains two intrachain disulfide bridges and is expressed as a 36 kDa noncovalently associated homodimer (6, 8, 9). The IL‑10 dimer binds to two IL‑10 R alpha /IL‑10 R1 chains, resulting in recruitment of two IL‑10 R beta /IL‑10 R2 chains and activation of a signaling cascade involving JAK1, TYK2, and STAT3 (10). IL‑10 R beta does not bind IL‑10 by itself but is required for signal transduction (1). IL‑10 R beta also associates with IL‑20 R alpha , IL‑22 R alpha , or IL‑28 R alpha to form the receptor complexes for IL‑22, IL‑26, IL‑28, and IL‑29 (11-13). IL‑10 is a critical molecule in the control of viral infections and allergic and autoimmune inflammation (14 ‑ 16). It promotes phagocytic uptake and Th2 responses but suppresses antigen presentation and Th1 proinflammatory responses (2).

1. Pestka, S. et al. (2004) Annu. Rev. Immunol. 22:929.
2. Sabat, R. et al. (2010) Cytokine Growth Factor Rev. 21:331.
3. Mathurin, P. et al. (2002) Am. J. Physiol. Gastrointest. Liver Physiol. 282:G981.
4. Grewe, M. et al. (1995) J. Invest. Dermatol. 104:3.
5. Szony, B.J. et al. (1999) Mol. Hum. Reprod. 5:1059.
6. Vieira, P. et al. (1991) Proc. Natl. Acad. Sci. 88:1172.
7. Hsu, D.-H. et al. (1990) Science 250:830.
8. Windsor, W.T. et al. (1993) Biochemistry 32:8807.
9. Syto, R. et al. (1998) Biochemistry 37:16943.
10. Kotenko, S.V. et al. (1997) EMBO J. 16:5894.
11. Kotenko, S.V. et al. (2000) J. Biol. Chem. 276:2725.
12. Hor, S. et al. (2004) J. Biol. Chem. 279:33343.
13. Sheppard, P. et al. (2003) Nat. Immunol. 4:63.
14. Fitzgerald, D.C. et al. (2007) Nat. Immunol. 8:1372.
15. Wu, K. et al. (2007) Cell. Mol. Immunol. 4:269.
16. Blackburn, S.D. and E.J. Wherry (2007)Trends Microbiol. 15:143.

Bioinformatics

• Gene Symbol: IL10
• Uniprot:
  • P22301.1()