The most prominent members of the interleukin-1 (IL-1) superfamily are IL-1alpha and IL-1beta. They lack a signal peptide and are secreted by an unconventional, endoplasmic reticulum-Golgi-independent mechanism. IL-1alpha was reported to be more widely and constitutively expressed and has intracellular functions, but also acts locally in a membrane-bound form by activating IL-1R1. Additionally, passive release of IL-1alpha upon cell death can trigger a sterile inflammatory response to dying cells. The cleavage of IL-1alpha is not mediated by caspase-1 and is not re- quired for binding to IL-1R1. Recently it has been observed that all activators of the inflammasome NLRP3/NALP3 induce the simultaneous secretion of IL-1alpha and IL-1beta. Although most activators fully rely on the inflammasome for IL-1alpha secretion, some induce the processing and secretion of IL-1alpha in an inflammasome-independent manner
Sample Type:Cell culture supernatants, serum and plasma
Intended Use:The mouse IL-1 alpha BioAssay™ ELISA Kit is to be used for the in vitro quantitative determination of mouse IL-1alpha in cell culture supernatants, serum and plasma.
Sensitivity:9pg/ml
Range:15.6pg/ml-1ng/ml
Specificity:Recognizes natural and recombinant mouse IL-1alpha (proform and p17 fragment). Does not detect human IL-1alpha or mouse IL-1beta.
Test Principle:This BioAssay™ is a sandwich Enzyme Linked-Immunosorbent Assay (ELISA) for quantitative determination of mouse IL-1alpha in cell culture supernatants, serum and plasma. A monoclonal antibody specific for IL-1alpha has been precoated onto the 96-well microtiter plate. Standards (STD) and samples are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, IL-1alpha is recognized by the addition of a biotinylated monoclonal antibody specific for IL-1alpha (DET). After removal of excess biotinylated antibody, streptavidine-peroxidase (STREP-HRP) is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450nm after acidification and is directly proportional to the concentration of IL-1alpha in the samples.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
