Protein blot (Western) hub: Difference between revisions
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==Overview== | ==Overview== | ||
A Western Blot allows for the semiquantitative determination of protein expression. | A Western Blot allows for the semiquantitative determination of protein expression. | ||
Revision as of 03:32, 15 October 2008
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.
