Re: [ccp4bb] sodium thiocyanate as cryoprotectant

[abhimanyu singh]

Hi Alex,

You may also think of gradually increasing PEG3350 concentration to around
40% which has worked pretty well in some of my cases.

Greetings,
Abhi



Abhimanyu Kumar Singh, Ph.D.
School of Biosciences
University of Kent
Canterbury, UK.

On 27 Sep 2016 23:43, "Alex Lee"  wrote:

> Hi CCP4bb members,
>
> I have a condition for my protein crystals at "0.2M sodium thiocyanate,
> 25% PEG3350, pH6.9".
>
> I'd like to try cryoprotectant with just higher concentration of sodium
> thiocyanate, but I have no google hits of the range of concentration I
> should use.
>
> I wonder anyone in this forum tried to use sodium thiocyanate as
> cryoprotectant?  What's the concentration range for this if used as
> cryoprotectant?
>
> Thanks.
>

Re: [ccp4bb] sodium thiocyanate as cryoprotectant

[Srivastava, Dhiraj]

Probably you can not use thiocyanate as cryoprotectant because its a chaotropic 
agent. Although its a very mild denaturant (destabilizer) but at high 
concentration, it can destroy your crystals. I have recently used 30 % sucrose 
for cryo protection of crystals growing under same condition and it worked 
beautifully. My crystals were cracking in glycerol.

Dhiraj

On Sep 27, 2016, at 5:43 PM, Alex Lee 
> wrote:

Hi CCP4bb members,

I have a condition for my protein crystals at "0.2M sodium thiocyanate, 25% 
PEG3350, pH6.9".

I'd like to try cryoprotectant with just higher concentration of sodium 
thiocyanate, but I have no google hits of the range of concentration I should 
use.

I wonder anyone in this forum tried to use sodium thiocyanate as 
cryoprotectant?  What's the concentration range for this if used as 
cryoprotectant?

Thanks.

[ccp4bb] sodium thiocyanate as cryoprotectant

[Alex Lee]

Hi CCP4bb members,

I have a condition for my protein crystals at "0.2M sodium thiocyanate, 25%
PEG3350, pH6.9".

I'd like to try cryoprotectant with just higher concentration of sodium
thiocyanate, but I have no google hits of the range of concentration I
should use.

I wonder anyone in this forum tried to use sodium thiocyanate as
cryoprotectant?  What's the concentration range for this if used as
cryoprotectant?

Thanks.

[ccp4bb] MXIS 2016

[ferrer]

The deadline for subscription is extended  to 7 Oct.
Still some places available

*



MXIS 2016: Practical course on in-plate crystallography, with special 
focus on ligand screening*



*Part 1: *7-9 Nov., at the CNRS campus, Montpellier, France

*Part 2: *30 Nov. - 2 Dec., at the EPN campus, Grenoble, France


*The MXIS training course is dedicated to in situ protein 
crystallography, with a special emphasis on ligand screening. This 
workshop is organized in two parts, each one lasting for 3 days. It 
includes:*


**

 *

   Part 1 (Montpellier): full lectures at the “Centre de Biochimie
   Structurale”followed by practicals. Topics: platepre-coating
   for“Dry” co-crystallization [1], in situ screening on a lab source).

 *

   Part 2 (Grenoble): full lectures at the “Institut de Biologie
   Structurale” and practical sessions at the ESRF, on beamlines
   FIP-BM30A and ID30B. Full classes will be done in day 1 while
   practical sessions will be organized in subgroups, from day 2 to day
   3. Topics: in situ diffraction for crystal screening and structure
   resolution, in situ data processing.

Possibility will be given to participants to practice on their own samples.

The workshop is limited to 6 and 8 participants for Part 1 and 2 
respectively.


No fees are required.

The application period will close on the 7th October. Applicants will be 
required to submit a short CV and a motivation letter to 
jean-luc.fer...@ibs.fr 


More information on https://workshops.ibs.fr/MXIS-2016

MXIS 2016 is made possible by the contributions of: IBS, CBS, EMBL, 
ESRF, FRISBI, ...


[1] Combining 'dry' co-crystallization and in situ diffraction to 
facilitate ligand screening by X-ray crystallography. Gelin M, Delfosse 
V, Allemand F, Hoh F, Sallaz-Damaz Y, Pirocchi M, Bourguet W, Ferrer JL, 
Labesse G, Guichou JF. Acta Crystallogr D Biol Crystallogr. 2015 
Aug;71(Pt 8):1777-87. doi: 10.1107/S1399004715010342.



--

Jean-Luc Ferrer
Institut de Biologie Structurale
71 Avenue des Martyrs
CS 10090
38044 Grenoble Cedex 9 - FRANCE
Ph.: +33 (0)4 57 42 85 22
Fax: +33 (0)4 76 50 63 14