Dear Reza,
in the past I had work with protein able to oligomerize reversibly but
when oligomerization happened, even if I was able to separate and obtain
monomeric protein,
protein was not in a good condition.
Have you try to characterize the two different states by DLS ? To
discriminate "interaction with resin" that you suspect from oligomerization.
Nicolas
Nicolas Foos
PhD
Structural Biology Group
European Synchrotron Radiation Facility (E.S.R.F)
71, avenue des Martyrs
CS 40220
38043 GRENOBLE Cedex 9
+33 (0)6 76 88 14 87
+33 (0)4 76 88 45 19
On 12/01/2017 01:50, Christopher Colbert wrote:
What's your monomeric molecular weight? Increased salt concentration
can easily drive oligomerization.
What is your evidence that it interacts with the resin?
Cheers,
Chris
--
Christopher L. Colbert, Ph.D.
Associate Professor
Department of Chemistry and Biochemistry
North Dakota State University
P.O. Box 6050 Dept. 2710
Fargo, ND 58108-6050
PH: (701) 231-7946
FAX: (701) 231-8324
From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK
<mailto:CCP4BB@JISCMAIL.AC.UK>> on behalf of Reza Khayat
<rkha...@ccny.cuny.edu <mailto:rkha...@ccny.cuny.edu>>
Reply-To: Reza Khayat <rkha...@ccny.cuny.edu
<mailto:rkha...@ccny.cuny.edu>>
Date: Wednesday, January 11, 2017 6:42 PM
To: "CCP4BB@JISCMAIL.AC.UK <mailto:CCP4BB@JISCMAIL.AC.UK>"
<CCP4BB@JISCMAIL.AC.UK <mailto:CCP4BB@JISCMAIL.AC.UK>>
Subject: Re: [ccp4bb] Off-topic question about SEC
All these make sense. Protein is very strange cause it goes from
60kDa (globular) to an apparent 360kDa. Process is reversible too.
Reza Khayat, PhD
Assistant Professor
City College of New York
Department of Chemistry
New York, NY 10031
------------------------------------------------------------------------
*From:* CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK
<mailto:CCP4BB@JISCMAIL.AC.UK>> on behalf of Keller, Jacob
<kell...@janelia.hhmi.org <mailto:kell...@janelia.hhmi.org>>
*Sent:* Wednesday, January 11, 2017 7:39 PM
*To:* CCP4BB@JISCMAIL.AC.UK <mailto:CCP4BB@JISCMAIL.AC.UK>
*Subject:* Re: [ccp4bb] Off-topic question about SEC
Yes if it either
A) oligomerizes
B) significantly changes shape
C) aggregates reversibly
On option B: Lower NaCl could make the protein “appear” bigger by
unfolding it a bit; hydrophobic interactions should be weaker in lower
NaCl.
JPK
Artem
www.harkerbio.com <http://www.harkerbio.com>
"where wild SEC columns roam free"
On Jan 11, 2017 7:22 PM, "Reza Khayat" <rkha...@ccny.cuny.edu
<mailto:rkha...@ccny.cuny.edu>> wrote:
Hi,
Sorry for the off-topic question. Can a protein in lower [NaC] run
faster on a SEC than at higher [NaCl] (i.e. elute at an earlier
volume)? The protein elutes well within the resolution limits of
the SEC with a symmetric gaussian A280 profile. I know that at
lower [NaCl] the protein can elute later because it may
interact with the matrix. Thanks.
Best wishes,
Reza
Reza Khayat, PhD
Assistant Professor
City College of New York
Department of Chemistry
New York, NY 10031
