Harsh,
This article describes common buffers for CD on page 2:
http://www.ncbi.nlm.nih.gov/pubmed/17406547
Or this article on page 8:
http://www.ncbi.nlm.nih.gov/pubmed/16027053
It seems like phosphate is the best, because it has low absorption at
180-200nm region. From organic buffers Tris/H2SO4
Hi,
Sorry for off-topic question.
Does anyone have experience of the stabilisation of water-soluble proteins
by detergents? Protein I'm working with is definitely water-soluble and has
high yield, but, unfortunately, not very stable. Especially during
concentration. So, we thought that adding som
Alexander,
We produce baculovirus at Gibco SF900-II + 10% heated FBS + 1xPSG and then
store at 4C routinely. And it seems to be quite stable. We have a virus
that had no change for more than 2 years: we judge by the volume of it
needed to infect_cells/produce_protein, no titer measurements. We als
of GSH?
>
> Thanks,
> Peter
>
>
> On Wed, 29 Aug 2012, Vitali Stanevich wrote:
>
> Peter,
>>
>> Such high concentration of GSH may change the pH according to our
>> experience. We usually use 50 mM Tris pH=8.0, 5 mM GSH, 3 mM DTT - so that
>> you can
Peter,
Such high concentration of GSH may change the pH according to our
experience. We usually use 50 mM Tris pH=8.0, 5 mM GSH, 3 mM DTT - so that
you can load the sample on ion-exchange column after elution. If your
protein is not stable without NaCl - you should add it also.
Vitali
On Wed, Au
The one we have is rubbish. I can tell a vendor name if you want off-list.
A lot of non-specific bands + high background level makes it almost
impossible to use against cell lysate. It sort of works against purified
His-tag protein, but not perfect either.
On Mon, Jun 25, 2012 at 4:33 PM, D Bonsor
Donghui,
There is detergent screen from Hampton:
http://hamptonresearch.com/product_detail.aspx?cid=1&sid=39&pid=31
We set up detergent screen for crystal conditions we'd like to improve:
200nl(buffer) + 200nl(protein) + 40nl(detergent). Occasionally (very
occasionally) it works, but don't put to
Gloria,
Why don't you introduce TEV-cleavage site? You can clone it between His-
and sumo-protein_of_interest (if you want co-crystallise sumo with your
protein) or between His-sumo and protein_of_interest (if you want to
crystallise your protein only).
Vitali
On Thu, May 24, 2012 at 2:41 PM, Gl
You can try this site:
http://espript.ibcp.fr/ESPript/cgi-bin/ESPript.cgi
If you have .pdb for one of your sequences, you'll be able to show secondary
structure also.
vitali
On Thu, Aug 18, 2011 at 6:31 PM, Yuri wrote:
> Hello Everyone,
> A little off topic but, what is a good way to show (pub
