Protein blot (Western) hub: Difference between revisions
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*[[Ketner: Tissue culture viral lysate Western Blot]] | *[[Ketner: Tissue culture viral lysate Western Blot]] | ||
*[[Western Blot/Tissue Preparation|Tissue preparation for western blot]] | *[[Western Blot/Tissue Preparation|Tissue preparation for western blot]] | ||
==External links== | |||
* [http://en.wikipedia.org/wiki/Western_blot Western blot on Wikipedia] | |||
* [http://www.protocol-online.org/biology-forums/SDS-PAGE-and-Western-Blotting/ Discussion on Western blots on Protocol Online] | |||
[[Category:Protocol]] | [[Category:Protocol]] | ||
[[Category:In vitro]] | [[Category:In vitro]] | ||
[[Category:Protein]] | [[Category:Protein]] |
Revision as of 03:35, 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
- Endy:Tris-Tricine Acrylamide Gels
- Endy:E. coli Western Blot
- Endy: General Western Blot
- Odom:Western Blot
- Sauer:Western blot
- Ketner: Tissue culture viral lysate Western Blot
- Tissue preparation for western blot