Coomassie Blue: destaining

Allan Stensballe allan at NUKESPAMMERSstensballe.com
Sun Jul 16 15:14:25 EST 2000


On Sun, 16 Jul 2000 11:17:44 -0400, pxpst2 at dlt.pitt.edu (Peter
Pediaditakis) wrote:

>> For visualization of ng levels of protein you should go for silver
>> staining instead of CB or Sypro dyes (assumed that you stain
>> electrophoretic gels and just need visualization).    
>> Silver staining with the right shortcuts is nearly as fast as CB, but
>> have sub ng sensitivity.
>
>Draw back of Silver is that you can not sequence the protein......but then
>again if one is looking at sequencing 10 ng of protein then they must be
>working with a god on the MS MS sequencer.

Well, I ment for _detection_ of gel bands of 10 ng protein. The
questioner did not specify the further use of the protein, but I
assumed that blotting/electroeluding the protein wasn´t the case ;-)  

Silver staining is a common staining technique prior to analysis by
mass spectrometry. Tandem MS sequencing of peptides generated from 10
ng protein (fmol range) from silver stained gels are routinely
performed in most labs specialized in biological MS (hybrid
instruments like nESI-QTOF/QqTOF have low(sub) fmol sensitivity). 

BTW: Dr. Martins might find this paper from Science's STKE
interesting.
http://www.stke.org/cgi/content/full/OC_sigtrans;2000/37/pl1
----------------------------------------------------
Allan Stensballe,
Protein Research Group,
University of Southern Denmark,
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