Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-10 has been pre-coated onto a microplate. Standard, control, or sample and the working solution of Biotin-Conjugate are pipetted into the wells. Following incubation and wash steps, any IL-10 present is bound by the immobilized antibody and the detection antibody specific for IL-10 is binds to the combination of capture antibody-IL-10 in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of IL-10 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-10 standard dilutions and IL-10 sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
Interleukin-10 (IL-10), also known as cytokine synthesis inhibitory factor (CSIF), is the charter member of the IL-10 α-helical cytokine family that also includes IL-19, IL-20, IL-22, IL-24, and IL-26/AK155. IL-10 is a pleiotropic cytokine that exerts immunosuppressive or immunostimulatory effects in a variety of cell types. In 1989, Fiorentino et al. found that the mouse Th2 cell line D10.G4.1 produced a new cytokine that inhibits the transcription of cytokine mRNA in Th1 cell line, called cytokine synthesis inhibitory factor (CSIF) and named IL-10 in the same year.
Mouse, rat and human IL-10 cDNA sequences each contain 178 amino acid residues and have an 18 amino acid signal peptide sequence. The mature IL-10 molecule is 160 amino acid residues, and the mouse and human IL-10 molecules contain 5 and 4 cysteine residues respectively. Their molecular weight ranges from 35-40 kDa. At the amino acid level, rat IL-10 has 85% and 74% homology to human and mouse IL-10 respectively. Rat and human IL-10 act on mouse-derived cells, whereas mouse IL-10 does not cross-react to human cells.
IL-10 mediates its biological activities through a heteromeric receptor complex composed of the type II cytokine receptor subunits IL-10 Rα and IL-10 Rβ. IL-10 Rα is a 110 kDa transmembrane glycoprotein that is expressed on lymphocytes, NK cells, macrophages, monocytes, astrocytes, intestinal epithelial cells, cytotrophoblasts, and activated hepatic stellate cells, while the 75 kDa transmembrane IL-10 Rβ is widely expressed. The IL-10 dimer binds to two IL-10 Rα chains, triggering recruitment of two IL-10 Rβ chains. IL-10 Rβ does not bind IL-10 directly but is required for signal transduction. IL-10 Rβ also associates with IL-20 Rα, IL-22 Rα1, or IL-28 Rα to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29.
Antagonists of IL-10 may have anti-EB virus effects; IL-10 may become an anti-inflammatory treatment by promoting the expression of IL-1 Rα by monocytes. Animal experiments show that IL-10 can effectively prevent the death of mouse shock induced by LPS.
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with antibody to rat IL-10 (8-12)||1 plate|
|Rat IL-10 standard lyophilized, 2000pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-rat IL-10 antibody||2 vials|
|Streptavidin-HRP solution||2 vials|
|standard /sample Diluent||1 bottle|
|Biotin-Conjugate antibody Diluent||1 bottle|
|Streptavidin-HRP Diluent||1 bottle|
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 vial|
|Stop Solution||1 vial|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 15pg/mL.|
|ASSAY RANGE||31.25 - 2000 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant Rat IL-10. The factors listed below were prepared at 50ng/ml in standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.|
Factors assayed for cross-reactivity
|Recombinant rat||CINC-1, GDNF, GM-CSF, IFN-γ, β-NGF, PDGF-BB, TNF-α, IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-18|
|Recombinant human||IL-10, IL-10sR|
|Recombinant mouse||IL-10 sR|
Data Analysis Assistance
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