IL-1 alpha and IL-1 beta are both synthesized as 31kD precursors that are subsequently cleaved into proteins with molecular weights of approximately 17,000. Neither precursor contains a typical hydrophobic signal peptide sequence and most of the precursor form of IL-1 alpha remains in the cytosol of cells，although there is evidence for a membrane-bound form of the precursor form of IL-1 alpha. The IL-1 alpha precursor reportedly shows full biological activity in the EL 4 assay. Among various species，the amino acid sequence of mature IL-1 alpha is conserved 60% to 70% and human IL-1 has been found to be biologically active on murine cell lines. Both forms of IL-1 bind to the same receptors，designated type I and type II. Recent evidence suggests that only the type I receptor is capable of signal transduction and that the type II receptor may function as a decoy，binding IL-1 and thus preventing binding of IL-1 to the type I receptor.Sample Type：Cell culture supernatants，serum，plasma，and tissueIntended Use：The Mouse IL-1alpha BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Mouse IL-1alpha concentrations in cell culture supernatants，serum，plasma，and tissue.Sensitivity：4pg/mlRange：7.8-500pg/mlSpecificity：Recognizes mouse IL-1alpha.Sample Volume：100ul/wellTest Principle：This assay employs the Sandwich immunoassay technique. An anti-m IL-1alpha monoclonal coating antibody is adsorbed onto microwells. IL-1alpha 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-1alpha antibody is added and binds to IL-1alpha captured by the first antibody. Following incubation unbound Biotinylated anti-m IL-1alpha antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-m IL-1alpha antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of IL-1alpha 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-1alpha standard dilutions and IL-1alpha sample concentration determined.Kit Components：Aluminium pouches with a Microwell Plate： 8x6Standard： 1xVialBiotinylated antibody： 1xVialStreptavidin-HRP： 1xVialAssay Buffer （25ml/Vial）： 1xVialWash Buffer Concentrate 20x （30ml/Vial）： 1xVialTMB Substrate Solution： 1x6mlStop Solution （12ml/Vial）： 1xVialPlate Covers- Adhesive strips： 2xstripsInstruction： 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.
