Hi Sebastiano,
Just to add to Dima's response, I've used ceramic hydroxyapatite column and
got great results separating 2 very large membrane proteins complexes that
eluted together in DEAE/Q and ran in similar size range in SEC. One eluted
in 100-200mM KPi and the the other in 300mM+ KPi ~pH 7.4.
Hi Krish,
Seconding Tim's comments, you may need to switch construct or expression
host. I've seen many times when a membrane protein seems to be in the yeast
membrane, but seems unextractable or most likely, improperly folded. Since
you're already working with yeast, I would recommend trying S.ce
orse in OG), and my solution
was to add more Ulp1 and it worked beautifully and I've gotten nice
crystals of my protein.
-john lee
On Sun, Feb 16, 2014 at 9:58 AM, Raji Edayathumangalam wrote:
> Hi Everyone,
>
> After several attempts to cleave the SUMO tag off my membrane
Hi Theresa,
I totally agree with the bead beater idea.
Good reference for recipe (pg.699-701):
Overexpression and Purification of Integral Membrane Proteins in Yeast
Franklin A. Hays,*,1 Zygy Roe-Zurz,† and Robert M. Stroud
-John
On Wed, Oct 8, 2014 at 7:25 AM, Theresa Hsu wrote:
> Dear
I have a structure of a transcription factor in which the recognition helix
from two different protomers bind to the same (stretched out) major groove.
I was curious whether anyone have seen this kind of motif before - two
anti-parallel helices in same major groove. I'm searching the pdb/google
bu
Hi everyone,
Slightly off topic here but I got myself volunteered by my 2nd grade son to
do a show and tell at his class. I have the rock candy experiment ready
with some background info on what I do.
Can anyone direct me to some resources or your personal demo's that you
have done?
Thanks a bun
Hi Rhys,
The bOG concentration of 1.0% is quite acceptable and if your protein is
stable and happy in OG, then you're in luck. At 30% PEG 600, I'm not
surprised you're seeing the phase separation or detergent 'blob's for lack
of a better term. I believe you have the latter, which many of us workin
