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ELISA


ELISA's provide a wealth of information in their simplests, but they can also be useed in more complex versions to provide enhanced signals, for more precise results.
Direct ELISA
An antigen coated to a multiwell plate is detected by an antibody that has been directly conjugated to an enzyme.
Indirect ELISA
Antigen coated to a polystyrene multiwell plate. First an unlabeled primary antibody, which is specific for the antigen, is applied. Next, an enzyme-labeled secondary antibody is bound to the first antibody. The secondary antibody is usually an anti-species antibody and is often polyclonal.

Sandwich ELISA
Sandwich ELISA's typically require matched antibody pairs, where each antibody is specific for a different, non-overlapping part (epitope), of the antigen molecule. The first antibody, termed the capture antibody, is coated to the polystrene plate. The analyte or sample solution is added to the well. A second antibody, the detection antibody, follows this step in order to visualize the amount of analyte figuur 5 present. Polyclonals can also be used for capture and/or detection in a sandwich ELISA, provided that they are different enough to allow both capture and detection of the analyte through different epitopes. If the detection antibody is conjugated to an enzyme, then the assay is called a direct sandwich ELISA. If the detection antibody is unlabeled, then a second detection antibody will be needed resulting in an indirect sandwich ELISA.


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