Is cell lysate in lysis buffer ok overnight?

Go to the profile of amarthaker
Aug 31, 2017

Can I keep protein extracted from E. coli bacteria (strain BL 21 DE3) in lysis buffer over night after centrifuging and filtering it?

Background: I am currently over expressing Maltose Binding Protein from BL21(DE3) strain of E.coli. For purification I pass it through maltose resin affinity column. Before that I add cell lysis buffer, sonicate it, centrifuge it to remove the precipitate and filter the supernatant with protein.

Problem: The procedure is long, and I go short in time on one day. Can I do all the procedure up to filtering the supernantant before the day of purification and keep the protein in cell lysis buffer over night for chromatography the next day?

Does it affect the protein in any way ?


Go to the profile of mdfenko
mdfenko 6 months ago

it really depends on the protein, storage conditions and the constituents of the lysis buffer.

Go to the profile of relaxin
relaxin 6 months ago

I would keep it frozen, and thaw it on ice before chromatography.