To determine certain infectious conditions.
The enzyme can be linked to a protein such as streptavidin if the primary antibody is biotin labeled. The most commonly used enzyme labels horseradish peroxidase (HRP) and alkaline phosphatase (AP). Other enzymes have been used as well, but they have not gained widespread acceptance because of limited substrate options. ELISA (enzyme-linked immunosorbent assay) is a plate-based assay technique designed for detecting and quantifying substances such as peptides, proteins, antibodies and hormones. Other names, such as enzyme immunoassay (EIA), are also used to describe the same technology. ELISA, or EIA, is an acronym for enzyme-linked immuno assay. ELISA is a test that detects and measures antibodies in your blood. This test can be used to determine if you have antibodies that are related to certain infectious conditions. ELISA testing is a double antibody “sandwich” immunoassay, which employs speci˜c antibodies against the disease to be tested: monoclonal antibody immobilized at the bottom of the microtiter wells, and polyclonal antibodies coupled with horseradish peroxidase as the conjugate solution. The resultant product, resorufin, is a highly colored and fluorescent compound. One commonly used enzyme conjugate in ELISA is horseradish peroxidase. Horseradish peroxidase (HRP) is a 40,000 Dalton protein, which catalyzes the reduction of hydrogen peroxide (H 2 O 2) to water (H 2 O) [1]. QUALITY ASSURANCE WHAT IS AN ELISA? ELISA is the abbreviation for enzyme-linked immunosorbent assay which is a diagnostic test employing the principle of double antigens “sandwich immunoassay” coupled with the enzymes horseradish peroxidase or alkaline phosphatase. The secondary antibody has specificity for the primary antibody. In a sandwich ELISA, it is critical that the secondary antibody be specific for the detection primary antibody only (and not the capture antibody) or the assay will not be specific for the antigen. Most ELISAs use antibodies conjugated to enzymes in order to generate signal through substrate reaction with the enzyme. HRP has been shown historically to be more sensitive than AP primarily due to its faster catalytic rate. However, HRP reactions may be self-limiting due to by-product inhibition of
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what is the purpose of peroxidase used in elisa