In the drop, the amSO4 becomes/casuses precipitate (I guess). The crystals were 
grown from the precipitate within a few days. When I looped up the crystal the 
precipitate was also looped up.

 

And I don't know whether AmSO4 is a cryoprotectant or not. When I freeze the 
crystal in LN, the crystal is not in a glassy drop under the microscope.

Rgds,

(HanJie)



 



Date: Fri, 3 Apr 2009 13:45:40 -0400
From: lambertus.vandenb...@umassmed.edu
Subject: Re: [ccp4bb] How to loop up crystal from AmSO4.
To: CCP4BB@JISCMAIL.AC.UK


I also don't understand what you mean by "The crystal always sticks with AmSO4" 
and "the crystal was still surrounded by AmSO4".... Is there solid AmSO4 in the 
drop? Judging from the concentration you state, this can't be the case.
Just loop up the crystal and freeze I would say, the MPD + PEG400 should be 
plenty high enough for good cryo.

Bert van den Berg
University of Massachusetts Medical School



-----Original Message-----
From: CCP4 bulletin board on behalf of James Holton
Sent: Fri 4/3/2009 1:36 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] How to loop up crystal from AmSO4.

If you want to get rid of mother liquid around crystal, mount it in oil.

Howerver, what is wrong with NH4SO4?  If your crystal can tolerate
near-saturated NH4SO4, it is a cryoprotectant all by itself.

-James Holton
MAD Scientist


HanJie_HCT Tai wrote:
> Hi,
> 
> I have a protein-DNA complex grown in 30 -40% MPD/0.2M Ammonium
> sulfate/5% PEG 400/ 0.1M Tris (8.5).
> 
> I believe that 30%MPD above does not require any cryoprotectant but i
> hv a problem to loop a crystal up. The crystal always sticks with AmSO4.
> 
> I tried to add 2ul up to 6ul reservoir solution into the protein drop
> (1+1 ul). Although AmSO4 was diluted, the crystal was still surrounded
> by AmSO4.
> 
> The loop size is 0.1 -0.2mm, the crystal is about 0.1 mm.
> 
> I tried to grow crystal without AMSO4 or 0.1M AmSO4, but no crystals
> was shown up.
> 
> Any suggestion to loop up the crystal without any AMSO4 attached?
>
> Regards,
> HCT (HanJie)
>
>
>
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