Artem,
Artem Evdokimov wrote:
Please note that osmotic shock extraction typically employs EDTA which is
obviously bad for IMAC.
This is not entirely correct.
I have used extracts with 5 mM EDTA for IMAC in the past.
If your IMAC column volume is large enough, only the top 1-2 mm will be
Sure, it's not always 'disastrously bad' to have EDTA (hence my use of the
word 'bad' rather than a more categorical statement. Donuts are bad for me
yet I can't stop eating them :)
Yes, you can take a risk. However since periplasmic isolation is already a
PITA, why add an extra concern?
Artem
Dear all,
I plan to prepare a protein fused to pelB signal peptide and secreted to
bacterial periplasmic space. Does anyone have a detailed protocol
for harvesting the protein from bacterial periplasmic space? The protein needs
to be in the native state and will be purified on a nickel column.
To: CCP4BB@JISCMAIL.AC.UK
Sent: Monday, February 09, 2009 12:52 PM
Subject: [ccp4bb] protocol for harvesting proteins from bacterial
periplasmic space
Dear all,
I plan to prepare a protein fused to pelB signal peptide and secreted to
bacterial periplasmic space. Does anyone have a detailed
-Original Message-
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Matt
Colins
Sent: Monday, February 09, 2009 12:53 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] protocol for harvesting proteins from bacterial
periplasmic space
Dear all,
I plan to prepare a protein fused
