Overview of Western Blot Hybridization

The term "blotting" refers to the transfer of biological samples from a gel to a membrane and their subsequent detection on the surface of the membrane. Western blotting (also called immunoblotting because an antibody is used to specifically detect its antigen) was introduced by Towbin, et al. in 1979 and is now a routine technique for protein analysis. The specificity of the antibody-antigen interaction enables a target protein to be identified in the midst of a complex protein mixture. Western blotting can produce qualitative and semiquantitative data about that protein.

Page Contents:

• Introduction
• Electrophoretic Separation of Proteins
• Transfer Proteins to a Membrane
• Blocking Nonspecific Sites
• Wash Buffer Formulations
• Primary and Secondary Antibodies
• Video: Probe the blot for your target proteins
• Detection Methods