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Determination of purity by SDS PAGE


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#1 Brightside

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Posted 14 April 2010 - 10:52 PM

Hi

We are trying to determine the purity of our proteins using SDS-PAGE. Is there a standard and accepted protocol that we can follow?
Specifically, we can effectively destain contaminating bands away if we destain for long enough. So, is there a specific length of time that we should destain for?

Any protocol or reference to an article would be highly appreciated!

Many thanks,

#2 K.B.

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Posted 15 April 2010 - 01:06 AM

Rule of thumb for destaining - "until background is clear".

#3 Inmost sun

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Posted 15 April 2010 - 12:31 PM

Hi

We are trying to determine the purity of our proteins using SDS-PAGE. Is there a standard and accepted protocol that we can follow?
Specifically, we can effectively destain contaminating bands away if we destain for long enough. So, is there a specific length of time that we should destain for?

Any protocol or reference to an article would be highly appreciated!

Many thanks,


after destaining the back ground, refer to the staining method; f.i. pure after silver staining;

if not pure, list the molecular masses of the contaminating bands




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