An indirect Enzyme-Linked Immunosorbent Assay (iELISA) is a common biochemical technique that detects an antibody using a labeled secondary antibody. This assay is primarily used to detect the total antibody amount in a sample. This method is commonly utilized to diagnose infection and to quantify antibodies against an invading antigen.
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RayBio iELISAs employ a specific antigen-coated 96-well plate. A sample containing the primary antibody is incubated into the wells and the antibodies present bind to the immobilized antigen. Next, the wells are washed and a biotin-labeled detection antibody that recognizes the primary antibody is added. After washing away unbound antibodies, an HRP-conjugated secondary antibody is then added, and when combined with a substrate, produces a signal amplification that is directly proportional to the amount of bound sample antibody.