Description
Interleukin-10 (IL-10), originally known as Cytokine Synthesis Inhibitory Factor (CSIF), is an 18.5 kD protein that shares over 80% sequence homology with the Epstein-Barr Virus protein BCRFI. The reported biological activities of IL-10, which may be interrelated, include inhibition of macrophage-mediated cytokine synthesis, suppression of the delayed type hypersensitivity response, and stimulation of the TH2 cell response, which results in elevated antibody production. The net effect of IL-10 action appears to be the inhibition of proinflammatory T cell mediated immunity. Recombinant human IL-10 (Cat No. 554611) is supplied as a frozen liquid comprised of 0.22 µm sterile-filtered aqueous buffered solution, containing glycerol and bovine serum albumin, with no preservatives. Recombinant human IL-10 is ≥ 95% pure as determined by SDS-PAGE analysis, and an absorbance assay based on the Beers-Lambert law. The endotoxin level is ≤ 0.1 ng per µg of human IL-10, as measured in a chromogenic LAL assay.
Preparation And Storage
Recommended Assay Procedures
Upon initial thawing the product should be aliquoted into polypropylene microtubes and frozen at -80°C for future use. Alternatively, the product can be diluted in sterile neutral buffer containing not less than 0.5 - 10 mg/mL carrier protein, such as human or bovine albumin, aliquoted and stored at -80°C. For in vitro biological assay use, carrier-protein concentrations of ≥ 1 mg/mL are recommended. For use as an ELISA standard carrier-protein concentrations of 5 - 10 mg/mL are recommended. Failure to add carrier protein or store at indicated temperatures may result in a loss of activity. The product should not be diluted to less than 50 μg/mL for long term storage. Carrier proteins should be pre-screened for possible effects in each investigator's experimental system. Carrier proteins may have an undesired influence on experimental results due to toxicity, high endotoxin levels or possible blocking activity.
ELISA Standard: Recombinant human IL-10 (Cat. No. 554611) is useful as a quantitative standard for measuring human IL-10 protein levels in an IL-10 specific sandwich ELISA with Purified Rat Anti-Human IL-10 (Cat. No. 554705) as a capture antibody and Biotin Anti-Human and Viral IL-10 (Cat. No. 554499) as the detection antibody. To obtain linear standard curves, investigators may want to consider using doubling dilutions of recombinant human IL-10 standard from 2,000 - 15 pg/mL to be included in each ELISA plate. For measuring human IL-10 in serum or plasma, investigators are highly encouraged to use the BD OptEIA™ Human IL-10 ELISA Set (Cat. No. 555157) and the BD OptEIA™ Human IL-10 ELISA Kit (Cat. No. 550613).
Bioassay: Investigators are advised that the Bioassay application is not routinely tested for this material and are highly encouraged to both titrate this material and include appropriate controls in relevant experiments. An activity range of 0.5 - 3.3 x 10^7 units/mg, encompassing an
ED50= 0.3 - 2 ng/mL, has previously been reported using MC/9 as indicator cells for proliferation, with a unit defined as the amount of material needed to stimulate a half-maximal response at cytokine saturation.
Blocking: Recombinant Human IL-10 (Cat. No. 554611) can be useful as a blocking control for flow cytometric analysis when used with fluorochrome conjugated antibodies, such as PE-conjugated JES3-9D7 (Cat. No. 554498). Investigators are advised that the blocking application is not routinely tested for this material. Intracellular cytokine staining techniques and use of blocking controls are described in detail by C.Prussin and D.Metcalfe.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Development References (3)
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Howard M, O'Garra A, Ishida H, de Waal Malefyt R, de Vries J. Biological properties of interleukin 10. J Clin Immunol. 1992; 12(4):239-247. (Biology). View Reference
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Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Biology). View Reference
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Vieira P, de Waal-Malefyt R, Dang MN, et al. Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: homology to Epstein-Barr virus open reading frame BCRFI. Proc Natl Acad Sci U S A. 1991; 88(4):1172-1176. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.