(on behalf of Jacob Keller
j-kell...@fsm.northwestern.edu)
Subject: [ccp4bb] SDS and IMAC
To: CCP4BB@JISCMAIL.AC.UK
Dear Crystallographers,
I am interested in doing a type of pull-down experiment by
immobilizing protein X on IMAC resin, flowing a large volume of dilute
lysate containing protein Y
Use Urea - it does not interfere with gels etc. Additionally, you should
consider covalent immobilization of protein X - using activated resins.
Amine and carboxylic acid immobilization is common, however my all-time
favorite is iodoacetamide-activated resin reacting with SH on the protein
(if you
Dear Crystallographers,
I am interested in doing a type of pull-down experiment by
immobilizing protein X on IMAC resin, flowing a large volume of dilute
lysate containing protein Y over it, then adding some concentrated
agent (solid SDS perhaps) to some more of the same lysate, and running
that
Hi Jacob
Why not try with urea and
for this type of studies I would probably use batch with the IMAC resin and not
run the samples over a column.
cheers
Preben
On 23/12/2010, at 16.11, Jacob Keller wrote:
Dear Crystallographers,
I am interested in doing a type of pull-down experiment
In my experience, either urea or guanidinium crashes out in gels. I
can't remember--which one is it? I am thinking guanidinium. (If the
answer to this email saves one grad student from the aggravation of
such a phenomenon, it will have been worth it...)
JPK
On Thu, Dec 23, 2010 at 9:31 AM,
In my experience, either urea or guanidinium crashes out in gels. I
can't remember--which one is it? I am thinking guanidinium. (If the
answer to this email saves one grad student from the aggravation of
such a phenomenon, it will have been worth it...)
It's GuHCl and what crashes is dodecyl
