Western Blotting Reagents

As with any immunostaining technique, Western blotting experiments contain multiple steps, each requiring various reagents, buffers, and solutions. Following protein gel electrophoresis, proteins must be transferred from an acrylamide gel onto a blotting membrane. This electrophoretic blot transfer calls for the use of transfer buffers. With proteins blotted on a membrane, antibody mixes, blocking solutions, and wash buffers need to be applied in order to detect specific protein targets. To visualize the antibody complexes, conjugated enzymes such as HRP or alkaline phosphatase will need to react with substrates to produce detectable signals. Other reagents may also be used if desired, such as signal enhancers or stripping buffers for removing antibodies. Here we have cataloged essential reagents used in Western blotting from a variety of scientific manufacturers.