IL-10 BioAssay™ ELISA Kit (Mouse) (Interleukin-10, IL-10, Cytokine synthesis inhibitory factor, CSIF, IL10)

Kits and Assays / Kits-ELISA (Cytokines, Interleukins), BioAssay™

4°C/-20°C

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-gamma) by activated Th1 cells, NK cells, and monocytes/macrophages. IL-10 has also been shown to inhibit macrophage cytotoxic activity and to stimulate the proliferation and differentiation of B cells, mast cells, and thymic T cells. 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 178aa residue precursor protein with a hydrophobic signal peptide that is cleaved to generate the 160aa 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.

Sample Type:
Cell culture supernatants, serum, plasma, and tissue

Intended Use:
The Mouse IL-10 BioAssay™ ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Mouse IL-10 concentrations in cell culture supernatants, serum, plasma, and tissue.

Sensitivity:
7pg/ml

Range:
15.625-1000pg/ml

Specificity:
Recognizes mouse IL-10. There was no detectable cross reactivity with any of the tested proteins following IL-1, 2, 3, 4, 5, 6, 8, 12, TNF, VEGF, TGF and IFN.

Sample Volume:
100ul/well

Test Principle:
This assay employs the Sandwich immunoassay technique. An anti-m IL-10 monoclonal coating antibody is adsorbed onto microwells. IL-10 present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-m IL-10 antibody is added and binds to IL-10 captured by the first antibody. Following incubation unbound Biotinylated anti-m IL-10 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-m IL-10 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored 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.

Kit Components:
Aluminum pouches with a Microwell Plate: 8x6
Standard: 1xVial
Biotinylated antibody: 1xVial
Streptavidin-HRP: 1xVial
Assay Buffer (25ml/Vial): 1xVial
Wash Buffer Concentrate 20x (30ml/Vial): 1xVial
TMB Substrate Solution: 1x6ml
Stop Solution (12ml/Vial): 1xVial
Plate Covers- Adhesive strips: 2xstrips
Instruction: 1x

Storage and Stability:
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.