I have collected digested protein for Salmonella and I'd like to have the peptides sequenced. Are there guidelines for determining the proper LC gradient time to separate peptides?
Dear Kevin Howe.,
Peptides can be separated by running a linear gradient of the organic solvent.
You should use RP-HPLC - LC-MS that means RP-HPLC connected with LC-MS, for better analysis of any proteins, Gradient time is 30mins, Gradient is
Starting 1%A & 99%B at 1st mins(Starting)
at 30mins(End) 100%A & 0%B at 30mins A-Water(TFA addition When required)
B-Acetonitrile , Then if you need more then 30mins for your doubt give to 45mins the gradient is same and give 214nm UV-Detection in the same time the LC-MS give report based on peaks appear.., Hope you understand.., if you want more details about instrumental methods just look my publications in my profile sections full-text free, and for your concern i have given following link...
Good Luck & Best Regards
R.Selva...........
http://systems.crump.ucla.edu/~sschen/JOCpaper.pdf
Exactly The R.Selva given details was give perfect result for you
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