Protein blot (Western) hub: Difference between revisions

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==Overview==
The '''protein blot''', also know as '''Western blot''' or '''immunoblot''', is one of the most tried and true experiments for [[Griffin:Antibody_Basics|antibody]]-dependent determination of protein expression/presence in a sample. This pages directs you both to the general and lab-specific protocols regarding protein blotting.
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 protein blot protocol==
* [[Western Blot (immunoblot)]]
* [[Protein blot (Western)]]


==Western blot protocols by individual labs==
==Western blot protocols by individual labs==
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==See also==
==See also==
* [[Griffin:Membrane_Stripping_and_Reprobing Membrane Stripping and Reprobing]]
* [[Griffin:Membrane Stripping and Reprobing]]
* [[Griffin:Western Blot Optimization]]


==External links==
==External links==
* [http://en.wikipedia.org/wiki/Western_blot Western blot on Wikipedia]
* [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]
* [http://www.protocol-online.org/biology-forums/SDS-PAGE-and-Western-Blotting/ Discussion on Western blots on Protocol Online]
* [[Image:3stars.png]] [http://www.lukemiller.org/journal/2007/08/quantifying-western-blots-without.html Quantifying protein blot (Western) band without expensive software - Photoshop, ImageJ]


[[Category:Protocol]]  
[[Category:Protocol]]  
[[Category:In vitro]]  
[[Category:In vitro]]  
[[Category:Protein]]
[[Category:Protein]]

Latest revision as of 02:17, 27 June 2012

The protein blot, also know as Western blot or immunoblot, is one of the most tried and true experiments for antibody-dependent determination of protein expression/presence in a sample. This pages directs you both to the general and lab-specific protocols regarding protein blotting.

General protein blot protocol

Western blot protocols by individual labs

See also

External links

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