Mouse IL-10 ELISA Kit

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. Standards and samples are pipetted into the wells and any IL-10 present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-10 is added to the wells and 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.

Target Information

Interleukin 10 (IL-10), initially designated cytokine synthesis inhibitor, is a pleiotropic cytokine that inhibits the production of a number of cytokines (including IL-1, GM-CSF, TNF, IL-6, IL-8, IL-12 and IFN-γ) by activated Th1 cells, NK cells, and monocytes/macrophages.

Mouse IL-10 is produced by a wide variety of cells, including activated Th2 cells, fetal thymocytes, monocytes/macrophages, keratinocytes, B cells, and glial cells. Mouse IL-10 cDNA encodes a 178 amino acid residue precursor protein with a hydrophobic signal peptide that is cleaved to generate the 160 amino acid residue mature protein. At the amino acid sequence level, there is approximately 73% identity between mouse and rat or mouse and human IL-10. Although human IL-10 is active on mouse cells, mouse IL-10 does not show species cross-reactivity on human cells. Herpes viruses, including EBV and equine herpes virus type 2, have been shown to encode viral homologues of IL-10 that exhibit some of the activities of IL-10 on both mouse and human cells.

IL-10 has been shown to inhibit macrophage cytotoxic activity and to stimulate the proliferation and differentiation of B cells, mast cells, and thymic T cells. IL-10 is a potent modulator of monocyte/macrophage function. IL-10 also enhances the release of soluble TNF receptors and inhibits the expression of surface ICAM-1 and B7. Finally, IL-10 has been reported to suppress the synthesis of superoxide anion plus reactive oxygen intermediates (ROI), and either inhibit or facilitate nitric oxide synthesis, depending on the time of exposure to activated macrophages. IL-10 has marked effects on B cells. IL-10 also has documented activity on endothelial cells, where it mimics IL-4, and on thymocytes and mast cells, where it acts as a growth costimulator.

GENE ID 16153

Materials Supplied

Kit Components 96 Wells Quantity/Size
Aluminium pouches with a Microwell Plate coated with monoclonal antibody to mouse IL-10(8﹡12) 1 plate
Mouse IL-10 Standard lyophilized, 1000 pg/ml upon reconstitution 2 vials
Concentrated Biotin-Conjugate anti-mouse IL-10 monoclonal 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

Performance Characteristics

REPEATABILITY The coefficient of variation of both intra-assay and inter-assay were less than 10%.
SENSITIVITY The minimum detectable dose was 7pg/mL.
ASSAY RANGE 15.6-1000 pg/mL
SPECIFICITY This assay recognizes both natural and recombinant mouse 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 human IL-10
Recombinant mouse IL-1α, IL-1β, IL-2, IL-3, IL-4
Recombinant porcine IL-10

Data Analysis Assistance

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