Re: [ccp4bb] sodium thiocyanate as cryoprotectant

[Alex Lee]

Hi All,

Thank you very much for the suggestions!

On Tue, Sep 27, 2016 at 4:37 PM, abhimanyu singh 
wrote:

> 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.
>>
>

[ccp4bb] AW: [ccp4bb] sodium thiocyanate as cryoprotectant

[Herman . Schreuder]

Dear Alex,

Did you try freezing with the mother liquor as is? If you freeze fast enough, 
you may not need additional cryoprotectant with 25% PEG3350. You should first 
freeze some reservoir solution and look if it stays clear, before attempting to 
destroy a crystal.

Best, Herman

Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] Im Auftrag von Alex Lee
Gesendet: Mittwoch, 28. September 2016 00:43
An: CCP4BB@JISCMAIL.AC.UK
Betreff: [ccp4bb] sodium thiocyanate as cryoprotectant

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

[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.