Molecular Biology

Experimental Protocol for Western Blotting

Western blot is a technique used to identify specific proteins from a complex mixture of proteins. Prior to Western blot, samples undergo a size-based protein separation step through gel electrophoresis. Western blot includes the transfer of size-based separated proteins onto a solid support and detection of specific proteins within the sample. Western blots are both highly sensitive and specific; the process incorporates a highly selective antibody detection system to identify target proteins within a sample which allows the researcher to accurately detect proteins based on the molecular weights. Individual steps in the procedure are modifiable and factors like gel concentration, duration of incubation, and voltage used in the run time can be optimized depending on the needs. These attributes make Western blot a highly accurate, flexible, and semi-quantitative method for identifying proteins.