Protein blot (Western) hub: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(see also)
Line 18: Line 18:


==See also==
==See also==
* [[Griffin:Membrane_Stripping_and_Reprobing Membrane Stripping and Reprobing]]
* [[Griffin:Membrane Stripping and Reprobing]]


==External links==
==External links==

Revision as of 06:18, 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.

General Western blot protocol

Western blot protocols by individual labs

See also

External links