What is Western Blot Protocol?
Western blot protocol is the exact standards by which the immunoblot test is conducted. Western blot is used to detect proteins in a tissue sample. The process separates native proteins by determining the lengths of polypeptides using a gel electrophoresis. The proteins themselves are then transferred to a nitrocellulose membrane and probed by antibodies. The first part of the western blot protocol begins with tissue collection. These samples are gathered either from living tissue or from a cell culture. Tissue is broken down and various inhibitors such as protease or phosphatase are introduced to prevent enzymes from performing digestion. This portion of the protocol is usually handled in cold temperatures to preserve the tissues. The gel electrophoresis process is the next step in western blot protocol. In this portion, the proteins are identified by various factors such as molecular weight or electrical charge. This process is most commonly completed using polyacrylamide gels with a
