Interferon gamma (IFN-gamma,also known as Type II interferon) was originally identified as an anti-viral activity produced by mitogen-activated T lymphocytes. The protein shares no significant homology with IFN-gamma or the various IFN-gamma family proteins. IFN-gamma is produced primarily by T lymphocytes and natural killer cells. The production of IFN-gamma is induced by antigenic challenge and by cytokines such as IL-12. Mouse IFN-gamma encodes a 155aa residue precursor protein with a hydrophobic signal peptide that is cleaved to generate the 133aa residue mature protein. In solution,IFN-gamma has been shown to exist as a non-covalently associated homodimer with topological similarity to IL-10. Mouse IFN-gamma shows approximately 40%aa sequence identity to human IFN-gamma and there is no cross-reactivity across species.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Mouse IFN-gamma BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Mouse IFN-gamma concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:7pg/mlRange:15.625-1000pg/mlSpecificity:Recognizes mouse IFN-gamma.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-m IFN-gamma monoclonal coating antibody is adsorbed onto microwells. IFN-gamma present in the sample or standard and Biotinylated anti IFN-gamma antibody binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard and Biotinylated anti IFN-gamma antibody are removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti IFN-gamma antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of IFN-gamma 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 IFN-gamma standard dilutions and IFN-gamma sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x12Standard: 2xVialBiotinylated antibody: 2xVialStreptavidin-HRP: 2xVialAssay Buffer (25ml/Vial): 2xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x12mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 3xstripsInstruction: 1xStorage 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.