Western blot, or western blotting, is a process that detects specific proteins in a complex mixture of proteins extracted from cells or tissues.
Western blot originated in the late 1970s and is now a routine protein analysis technique used extensively by biochemists, molecular biologists and cell biologists. The method results in both qualitative and semi-qualitative data and is helpful in a wide range of applications, including:
- Lyme disease ·
- BSE (Bovine Spongiform Encephalopathy)
The three main steps in the workflow are separation by size, transfer to stable support and visualization using appropriate primary and secondary antibodies.
Step one: Separation by size
Scientists and technicians use protein gel electrophoresis to separate protein mixtures according to the molecular weight of each component protein.
Step two: Transfer to stable support
After the proteins have been separated, they are transferred to a stable support membrane using a blotting apparatus.
Step three: Visualization with primary and secondary antibodies
In step three, antibodies specific to the protein of interest bind and are detected after the film is developed or digital images are processed.
