Protein blot (Western) hub

Overview

A Western Blot allows for the semiquantitative determination of protein expression. Crude cell lysates are loaded into a polyacrylamide gel containing a denaturing agent which give all the proteins a net negative charge. A current can then be passed through the gel and the proteins will migrate through the gel, with the largest proteins traveling the slowest, resulting in a lane where proteins become separated on the basis of their weight. The proteins can then be transferred from the gel onto a membrane (often nitrocellulose or PVDF), which can then by incubated with an antibody directed against the protein of interest. By using a detector conjugated to an antibody one can now detect specifically the protein of interest.

Relevant lab protocols

• Endy:Tris-Tricine Acrylamide Gels
• Endy:E. coli Western Blot
• Endy: General Western Blot
• Odom:Western Blot
• Sauer:Western blot
• Ketner: Tissue culture viral lysate Western Blot
• Tissue preparation for western blot

External links

• Western blot on Wikipedia
• Discussion on Western blots on Protocol Online