Description
Principle of the Assay: The LumiAb Human IL-1alpha Chemiluminescent ELISA Kit contains the components necessary for quantitative determination of natural or recombinant Human IL-1alpha concentrations within any experimental sample including cell lysates, serum and plasma. This particular immunoassay utilizes the quantitative technique of a "Sandwich" Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a "sandwich" format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on Human IL-1alpha while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and "sandwiching" of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a sensitive luminescent reaction to ensue upon substrate addition. When the Peroxide Enhancer solution is added, the reaction catalyzed by peroxidase yields light that is representative of the antigen concentration. After a brief incubation, the microplate can be read with a luminometer, allowing for generation of a standard curve and subsequent determination of protein concentration: