trypsin digestion

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trypsin digestion

nithya-5
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

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Re: trypsin digestion

Kyle Legate
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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