Dear all:
Any ideas to decrease protein polymer formation? my protein was easy to form
oligomers and precipitation when do purification,I have tried add glycerol and
DTT,both didn't work. Does anyone has experience to avoid it happening. Thanks!
Best Regards
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I am not sure what you mean by polymer formation. Presuming that you have
optimized your protein concentration, pH and salt concentration, you could
try arginine as an anti-aggregation agent in your purification (I presume
you do FPLC). Have a look at chaotropic agents used in protein
purification,
Keep the protein concentration low during purification steps along with using
other anti-aggregation agent/s. Make sure that the pH at which you are
purifying is not close to the pI of the protein. Until completely purified, all
purification steps should be performed in a cold room if it is a so
It really depends from the nature of the protein and if is
oligomerizing/agregating/forming polymers if any of this is reversible.
On the other side if you are working with one of the fibril forming
protein it will require optimizaiton on its on as they will form
naturally polymers.
Do you obs
It’s with much sadness that I learn of Ward’s passing.
To add to Bob’s message: Ward was elected as Chair of the IMCA’s supervisory
Board. He lead us through some difficult decisions, and always with a great
sense of humor
Thierry
> On Jul 18, 2020, at 7:36 AM, Sweet, Robert
> <27e0eb9d2
Hi Digant,
As Artem mentioned, it's a fairly simple process and it's similar to
plasmid transformation in E. coli. We work extensively with insect cells in
our lab, though we have never used electroporation, the normal chemical
transformation works quite well. Although bacmid requires careful handl
Hi Fellows,
Q for the Refmac cognosci:
Just curious: If I run unrestrained anisotropic refi on 1 A data, some
barely supported ligands display insane and physically impossible ADPs,
worthy of the ORTEP of the century. As cigars penetrate discs, it seems that
Hirschfeld criteria on
aniso A
Ward was the Program Official for all my grants! My life will not be
the same without him. He was always so supportive and helpful with
advice on how to navigate the sometimes convoluted system that is the
NIH. From the Protein Structure Initiative to today his hard work has
made possible my
Dear:
The protein was purified in 4 degree, and the expression level is low, so the
aggregation is not by high concentration; the buffer pH is 7.5 which is not
colse to the PI 8.6. It should be a dimer when function, but it was aggregated
when negative staining. Maybe I could try to add arginin
