Protein blot (Western) hub

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(added Colin North's overview on western blots)
Line 1: Line 1:
 +
==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==
==Relevant lab protocols==
*[[Endy:Tris-Tricine Acrylamide Gels]]
*[[Endy:Tris-Tricine Acrylamide Gels]]
*[[Endy:E. coli Western Blot]]  
*[[Endy:E. coli Western Blot]]  
*[[Endy: General Western Blot]]
*[[Endy: General Western Blot]]

Revision as of 18:55, 31 August 2006

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