ELISA (enzyme-linked immunosorbent assay) is a plate-based assay technique designed for detecting and quantifying peptides, proteins, antibodies and hormones. In an ELISA, an antigen must be immobilized to a solid surface and then complexed with an antibody that is linked to an enzyme. Detection is accomplished by assessing the conjugated enzyme activity via incubation with a substrate to produce a measureable product. The most crucial element of the detection strategy is a highly specific antibody-antigen interaction.
ELISAs are typically performed in 96-well (or 384-well) polystyrene plates, which will passively bind antibodies and proteins. It is this binding and immobilization of reagents that makes ELISAs so easy to design and perform. Having the reactants of the ELISA immobilized to the microplate surface makes it easy to separate bound from non-bound material during the assay. This ability to wash away nonspecifically bound materials makes the ELISA a powerful tool for measuring specific analytes within a crude preparation.
ELISA Technical Resources Center
Western blotting, ELISAs, and IHC are all immunoassays, and so all share in common the same basic…
ELISA Sample Preparation
Every immunoassay protocol begins with sample preparation. Western blot, IHC, and ELISA all require…
There are five types of ELISA, thus, about ELISA protocol, a few differences exist….
Though without that complicated protocols as IHC, immunoprecipitation etc,…
This part is specially intended to provide some tips for ELISA…