[Protein-analysis] Re: trypsin digestion

Kyle Legate legatek at hotmail.com
Sat Dec 17 13:07:39 EST 2005

nithya wrote:
> I have done a trypsin digestion of a coloured protein.........
> my protocol is 1mg/ml trypsin in 0.1% acetic acid.......
> protein soln 40 ug/ml in 100 mM AmBic buffer ph:8.5
> overnight incubation
> prot: trypsin ratio :  100:1/ 50:1
> my boss says that i need to c a reduction in color of the prot if
> digestion has occurred.....
> is ther any other way to confirm that digestion has occurred
> completely?
> I would welcome any suggestions on the protocol as well
> have to do MS of the digested peptides
> help me
> thanks
> Nithya
Run an SDS-PAGE gel--does your full-length protein disappear? You'll 
have to run a gel before MS anyway.

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