ginal Message -
From: "Katherine Sippel"
To: CCP4BB@JISCMAIL.AC.UK
Sent: Thursday, January 12, 2012 4:01:10 PM GMT -08:00 US/Canada Pacific
Subject: [ccp4bb] Metal won't strip from IMAC
Hi all,
I've run into a bit of a protein purification conundrum and wondered if anyone
had enc
Hi Katherine,
I recommend Zn-IDA Sepharose (Chelating Sepharose Fast Flow, GE Healthcare),
which we have been using successfully for more than 20 years, since the early
days of IMAC:
Skerra et al. (1991) The functional expression of antibody Fv fragments in
Escherichia coli: improved vectors a
Hi,
This is a fairly familiar issue. A hydrophobic protein with thiols often
contaminates IMAC columns and this kind of contamination can be difficult
to remove completely. In the past some of the methods that worked included
trypsin or papain treatment, followed by massive amounts of chelating
ag
uot;Katherine Sippel"
To: CCP4BB@JISCMAIL.AC.UK
Sent: Thursday, January 12, 2012 4:01:10 PM GMT -08:00 US/Canada Pacific
Subject: [ccp4bb] Metal won't strip from IMAC
Hi all,
I've run into a bit of a protein purification conundrum and wondered if anyone
had encountered a similar situa
Hi all,
I've run into a bit of a protein purification conundrum and wondered if
anyone had encountered a similar situation. I've exercised all of my
google-fu and can't find anything. It's a fairly straightforward setup;
His-tagged protein and Talon Co2+ resin, load lysate, wash with 5 mM
imidazol
