Western blotting is one of the most commonly used biochemical techniques to detect a specific protein from a mixture of proteins such as tissue extracts. Antibodies to the specific antigen are used to detect the protein. The mixture of proteins is resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a membrane. A specific antigen immobilized on the membrane is detected and visualized by a primary antibody, a secondary antibody-peroxidase conjugate, and a chemiluminescent reagent.
Key WordsWestern blotting SDS-PAGE primary antibody secondary antibody chemiluminescence