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Scientific method
 
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About Western transfer; after separating proteins by SDS-PAGE, why transfer the proteins to a nitrocellulose filter? Why not probe for the protein with antybody directly in the gel? Also why does the transfer have to be conducted immediately?

By singularitygirl Asked Oct 27 2009 10:48PM
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by Palifox on Nov 19, 2009 at 11:23 pm Permalink

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I'm not a biochemist but I have heard of this and checked it out a few days ago, then forgot your question. Apparently probing the gel directly does not work because the antibodies, being large molecules can't get at the analyte proteins which are buried in the gel. They analytes are moved from the gel onto the nitrocellulose (or PVDF) by capilliary action or "sideways" electrophoresis.

http://en.wikipedia.org/wiki/Western_blot
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About Western transfer; after separating proteins by SDS-PAGE, why transfer the proteins to a nitrocellulose filter? Why not probe for the protein with antybody directly in the gel? Also why does the transfer have to be conducted immediately?

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