Some Minor Questions Re SDS PAGE :

Chris LaRosa clarosa at biocomp.unl.edu
Mon May 8 10:05:03 EST 2000



> 1.  Should the protein standards be diluted with loading buffer before
> loading or     it is not needed?
> 
>It depends on what stds your using.   Read the manufacturers recomendations..
 2.  Do the protein standards also need to be boiled before loading?

Yes.

> 3.  Once boiled for 5 min with loading buffer, the protein stock samples
> could be stored in -20 but do they need to be boiled again each time before loading?
> Yes. 
> 4.  When I have far different concentrations of proteins, do I need to
> precipitate and dilute again in order to have approximately similar
> concentrations within equal total volume, in order to have a perfect run?  
Ideally yes... but often you can measure the protein concentrations and
adjust the volumes accordingly.  It helps to adjust each sample so that
you have equal volumes of sample buffer.




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