Description
Principle of the Assay: The LumiAb Human Gas6 Chemiluminescent ELISA Kit contains the components necessary for quantitative determination of natural or recombinant Human Gas6 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 Gas6 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: