Western Blotting

Western blotting, also known as immunoblotting, is a widely-used technique that utilizes antibodies for the detection of a specific protein within a sample. The process involves separating protein samples by electrophoresis and then transferring to a membrane, typically nitrocellulose or PVDF. Detection often involves the use of primary and secondary antibodies where the secondary antibody is conjugated either to an enzyme, such as HRP or AP, or to a fluorescent dye. The target protein can then be visualized using chemiluminescent or chemifluorescent detection reagents, or directly using the fluorescent tag. When purchasing supplies for Western blotting, the type of sample to be analyzed should be taken into consideration. Requirements for a small, relatively pure sample will be very different than those for a crude lysate or other complex mixture.