Protein blot (Western) hub: Difference between revisions
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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. | 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 (immunoblot)]] | |||
==Western blot protocols by individual labs== | |||
*[[Endy:Tris-Tricine Acrylamide Gels]] | *[[Endy:Tris-Tricine Acrylamide Gels]] | ||
*[[Endy:E. coli Western Blot]] | *[[Endy:E. coli Western Blot]] | ||
Revision as of 03:39, 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
- 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
