lower detection limit with Coomssie?

Troels Wind wind at biobase.dk
Fri Aug 30 02:17:23 EST 1996


Bruce W. Ritchings (bwr at icbr.ifas.ufl.edu) wrote:
: Hello Folks,
: 	For DNA work, the limit of visibility of a DNA band in an
: agarose gel is known to be around 10-15 ng.There are lots of
: variables, but that's the figure that's usually given (see Messing's
: primer on molecular biology, some years back). Is there a standard
: figure given for the limits of visibility for Coomassie-stained
: proteins in an SDS-PAGE gel? Any help greatly appreciated, thanks,
: Bruce Ritchings, University of Florida

Visibility in Coomassie-staining depends on the method used, i.e. 
quality of stain-solution and incubation procedures.

For a 'standard' procedure (staining with 0.25% R-250, 50% MeOH, 
10% HAc) 0.1-0.5 ug is visible.

If you want higher sensitivity, some modifications can enable you 
to see as little as 0.05 ug. Let me know if you want the protocol.

Cheers,
Troels



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