Hi allI work on an uropathogenic strain of E.coli. When I artificially express from a plasmid one of the transcriptional regulator, it results in secretion of a protein factor in the media, which has interesting biological roles. My aim is to purify this unknown factor present in the growth media and to identify it by Mass Spec.
But the problem is I am not familiar with the fractionation methods. Looking through the literature, I could see that many different protocols are being employed out of which I don’t know which the best is. I like your suggestion with regard to a good standard protocol for identifying an unknown protein in the growth media.
Again to put it differently, I have protein in the supernatant of the growth media. I just need to identify it.
Thanks for all those who read this and Thank you very much for someone who comments.