Protein blot (Western) hub

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
m (Western blot moved to Western blot (immunoblot) hub: longer Western blot page already exist; made this duplicate page an overview page linking to main WB page)
(illustration)
Line 1: Line 1:
 +
[[Image:Hub icon.png|right]]
 +
==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 06:32, 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.

Relevant lab protocols

Personal tools