Dear Ilme,
I am actually in your time zone -- Amst. NL. Wish I were qualified. I'll send
this to Marie.
Love Bob
On Oct 8, 2014, at 9:12 PM, Ilme Schlichting
wrote:
> The Project:
> X-ray free-electron lasers (XFELs) are poised to revolutionize structural
> biology. They provide highly int
The Project:
X-ray free-electron lasers (XFELs) are poised to revolutionize
structural biology. They provide highly intense, coherent, femtosecond
X-ray pulses that promise novel approaches to structure determination of
biological objects. We are at the forefront of this development,
designing
Dear Manjula Ramu,
You might want to consider immobilizing your thrombin-containing mixture
on a matrix (e.g. Bio-Rad's Affi-Gel, but there are many alternatives) and
then use the slurry for cleaving your recombinant protein. That way, you
never contaminate your sample with either thrombin or B
Hi Manjalu,
In my opinion, one way is to use iEX column. Depending of the Pi of you protein
and the one of the BSA, if they are sufficiently different, Ion exchange is on
possible way.
To separate your protein from BSA, you have to give us more information about
your protein if it's possible.
Dear Manjula,
Unless Thrombin requires BSA for functioning (I never used Thrombin) you
could run a size exclusion column with the Thrombin solution prior to
using it. Or you could by it from a different vendor.
Regards,
Tim
On 10/08/2014 03:27 PM, Manjula Ramu wrote:
> Hi all,
>
> My protein si
Hi all,
My protein size is nearly the same size of BSA and it has His tag. If I
have to cleave the tag I have to use Thrombin which is having huge amount
of BSA in it. Now how will I separate this BSA from my protein.
Thanks and Regards,
Manjula R
Research Scholar
Department of Biophysics
Nationa
Hi,
I would stay away from the Mem-PER kit since the detergents in there are
possibly quite harsh (?). I doubt that detergents would cause lysis of yeast
cell walls in any case.
There is a budget-friendly alternative for mechanical breaking of yeast and
that is the bead beater, sold by biospec.
Dear all
Would you have a good protocol for membrane protein extract from S. cerevisiae?
We don't have French press in our lab. Is the Pierce Mem-PER reagent kit useful
for proteins (10-12 transmembrane helices) that will be use for crystallization?
Thank you.
Theresa
Research Associate/Assistant with Murray Stewart at the MRC Laboratory
of Molecular Biology, Cambridge, UK.
It is anticipated that a Research Associate/Assistant position will soon
become available in Murray Stewart's group at the MRC Laboratory of
Molecular Biology in Cambridge to work on the
