Is cell lysate in lysis buffer ok overnight?

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Is cell lysate in lysis buffer ok overnight?

Postby amarthaker » Aug 31 2017 6:26 pm

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 ?
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Re: Is cell lysate in lysis buffer ok overnight?

Postby mdfenko » Sep 01 2017 11:40 am

it really depends on the protein, storage conditions and the constituents of the lysis buffer.
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Re: Is cell lysate in lysis buffer ok overnight?

Postby relaxin » Sep 02 2017 8:41 pm

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