Final answer:
Elisa and western blotting are immunoblot assays used to detect proteins, but they differ in technique and sensitivity. Elisa immobilizes antigens in a well and uses an enzyme-conjugated antibody to detect their presence. Western blotting separates proteins on a membrane and uses antibodies and enzymes or fluorophores to identify specific proteins.
Step-by-step explanation:
Elisa and western blotting are both immunoblot assays commonly used in molecular biology research to detect and identify specific proteins. However, they differ in their techniques and applications.
In Elisa, antigens are immobilized in a microtiter plate well. A specific antibody conjugated to an enzyme is added, and if the antigen is present, the antibody will bind. A colored end product is then generated to indicate the presence of the antigen. Elisa allows for fast detection, but it has lower sensitivity compared to western blotting.
In western blotting, proteins are separated by gel electrophoresis and transferred to a nitrocellulose membrane. A primary antibody, specific to the protein of interest, and a secondary antibody equipped with an enzyme or a fluorophore are added. The enzyme converts a colorless substrate into a colored product, or the fluorophore fluoresces. Western blotting is more sensitive than Elisa and can provide more detailed information about protein size and interactions.
ELISA is an immunosorbent assay that is used to detect antibodies or antigen in a sample. Western blotting is an analytical technique that is used to separate and identify proteins from a mixture.