Protein blot (Western) hub

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
m (See also)
(exchanged short intro from Western Blot (immunoblot) with long one from Western blot (immunoblot) hub)
Line 1: Line 1:
[[Image:Hub icon.png|right]]
[[Image:Hub icon.png|right]]
-
==Overview==
+
The Western blot is one of the most tried and true experiments for [http://openwetware.org/wiki/Griffin:Antibody_Basics antibody]-dependent determination of protein expression/presence in a sample.
-
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==
==General Western blot protocol==

Revision as of 08:21, 15 October 2008

The Western blot is one of the most tried and true experiments for antibody-dependent determination of protein expression/presence in a sample.

Contents

General Western blot protocol

Western blot protocols by individual labs

See also

External links

Personal tools