oard (on behalf of Jacob Keller
>)
>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
>l
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
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 su
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, Preben
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 experimen
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 o
