In-gel digestion using both Glu-C and trypsin

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pdow01
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In-gel digestion using both Glu-C and trypsin

Postby pdow01 » Thu Aug 07, 2014 12:54 am

Would anyone have a protocol that would allow me to perform a double digestion of gel bands using a combination of both Glu-C and trypsin?

Thanks,

Paul.

cavaliersun2003
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Postby cavaliersun2003 » Thu Nov 13, 2014 12:37 am

I never saw a protocol that use Glu-C and trypsin together. Since both work in pH8, so I guess you can do the digestion together in buffer like 0.1 M ammonium bicarbonate.
But I'm wondering what's the purpose is this for.....if you want to use multiple enzyme digestion just to get better coverage of your protein sequence in MS experiment, then I think it is better to do Glu-C and trypsin digestion in parallel, and run the LC-MS separately, then combined the results. I think digestion by both Glu-C and trypsin will make the peptide fragments too small.....


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