Hi!
I have some peptides that unfortunately have many products cracked in the spectrum (HPLC) close or combined at the peak of the chromatogram. I believe they are residues that came together in the cleavage. I separated the peptides in ice-cold ether but many did not precipitate, even with centrifugation they still carry these cleavage products. There is some protocol to better separate crude peptides from other products and achieve better separation for purification.
Thank you all
As you said cleavage products are interfering with peptide during observation, it can be assumed that they can be of smaller size residues in comparison to your target peptides than size exclusive chromatography might help here for separation.
если вы знаете размер определяемы пептидов, можно воспользоваться мембранными фильтрами на нужное количество дальтон. низкомолекулярные пройдут а то что выше останется. вот этот остаток и смыть с фильтра
You can try to optimize your protein precipitation step to reduce the noise (change solvent, volume, pH...), and/or combine the precipitation to a solid phase extraction and take advantage of the properties of your peptide to wash away the unwanted residues.
It really depends on the peptide you're working with.
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