Dear all,
I am expressing a 6xHis tagged secreted protein in a fermentor in P. pastoris,
using the standard minimal medium described in the invitrogen manual (plus
PTM1). Following collection of the culture medium, I am having problems with
purification of the protein as only a small fraction
College of Life SciencesBeijing, China
Date: Mon, 26 Mar 2012 15:04:43 +0300
From: peg...@pasteur.gr
Subject: [ccp4bb] recommendations_on_purification
To: CCP4BB@JISCMAIL.AC.UK
Dear all,
I am expressing a 6xHis tagged secreted protein in a fermentor in P. pastoris,
using the standard minimal
LaboratoryHarvard
Medical SchoolDana-Farber Cancer InstituteBoston, MA Peking UniversityThe
College of Life SciencesBeijing, China
Date: Mon, 26 Mar 2012 15:04:43 +0300
From: peg...@pasteur.gr
Subject: [ccp4bb] recommendations_on_purification
To: CCP4BB@JISCMAIL.AC.UK
Dear all,
I am
.
Postdoctoral Scientist, Wang Laboratory
Harvard Medical School
Dana-Farber Cancer Institute
Boston, MA
Peking University
The College of Life Sciences
Beijing, China
Date: Mon, 26 Mar 2012 15:04:43 +0300
From: peg...@pasteur.gr
Subject: [ccp4bb] recommendations_on_purification
To: CCP4BB
Hi Petros,
It's possible but unlikely that your media contains enough metal ions
to matter. More likely options include protein issues like
overgkycosylation, proteolysis (removal of the tag) or severe
aggregation. Since you are not experiencing this in BMGY, I wonder why
not to supplement your
I suspect that sometimes the protein chaperones the tag, which is solvent
exposed some fraction of the time. Try very slow loading or batch binding.
Kendall Nettles
On Mar 26, 2012, at 8:15 AM, Petros Giastas
peg...@pasteur.grmailto:peg...@pasteur.gr wrote:
Dear all,
I am expressing a
