Recombinant Human IL-10, Animal Free Protein

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Catalog # Availability Size / Price Qty
AFL1064-050
AFL1064-01M
Recombinant Human IL-10, Animal Free Protein Bioactivity
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Recombinant Human IL-10, Animal Free Protein Summary

Learn more about Animal-Free Recombinant Proteins

Product Specifications

Purity
>97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<0.10 EU per 1 μg of the protein by the LAL method.
Activity
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 #
N-terminal Sequence
Analysis
Met
Structure / Form
Noncovalently-linked homodimer
Predicted Molecular Mass
19 kDa
SDS-PAGE
18 kDa, reducing conditions

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AFL1064

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AFL1064

Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 100-500 μg/mL in PBS.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

Scientific Data

Bioactivity View Larger

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 0.0750-0.750 ng/mL. Two independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of Animal-Free™ IL-10.

SDS-PAGE View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IL-10

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).

References
  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.
Long Name
Interleukin 10
Entrez Gene IDs
3586 (Human); 16153 (Mouse); 25325 (Rat); 397106 (Porcine); 403628 (Canine); 102133450 (Cynomolgus Monkey); 493683 (Feline); 100135553 (Guinea Pig); 2949786 (Viral)
Alternate Names
CSIF; CSIFMGC126450; Cytokine synthesis inhibitory factor; GVHDS; IL10; IL-10; IL10A; IL-10MGC126451; interleukin 10; interleukin-10; TGIF

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.  

 Production

  • All molecular biology procedures use animal-free media and dedicated labware.
  • Dedicated fermentors are utilized in committed animal-free areas.

Purification

  • Protein purification columns are animal-free.
  • Bulk proteins are filtered using animal-free filters.
  • Purified proteins are stored in animal-free containers in a dedicated cold storage room.

    Quality Assurance

    • Low Endotoxin Level.
    • No impairment of biological activity.
    • High quality product obtained under stringent conditions.
    • For ex vivo research or bioproduction, additional documentation can be provided.

    Please read our complete Animal-Free Statement

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