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 tissue
Intended 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/ml
Range:7.8-500pg/ml
Specificity:Recognizes mouse IL-1alpha.
Sample Volume:100ul/well
Test 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.
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.
