I have the following proteins in the mentioned formulation:
pERK: 50 mM Tris-HCl, 150 mM NaCl, 0.25 mM DTT, 10 mM Glutathione, 0.1 mM EDTA, 0.1 mM PMSF and 25% Glycerol.
PPARg: 50% Glycerol, 20 mM Tris, pH 8
I have tried to do digestion following this protocol:
https://drive.google.com/open?id=0BwhAodGtCkmGT0JYTXozcVpQazA
But on LC-MS (ESI) I do not see any digested peptide. Any suggestion on what is going wrong here? Thanks.
Hi Ekram, your protocol seems solid, I would try to run it from step 10) with a standard peptide mix (i.e. which you also use for ms-calibration) and narrow down if there is a problem with the digest part or the zip-tip part of the protocol. Good luck!
Hallo Ekram, could you please see the attached protocol? i used it in my work.
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