Cryo
On 5 May 2015 13:51, faisaltari...@gmail.com wrote:
Thank you everyone..I have got some hints from this discussion and will
definitely try some of them to check its efficacy for my case...Thanx again
for your valuable suggestions..
On 5 May 2015 13:48, Anthony Savill
Hi Ursula,
you could try Perfluoropolyether PFO-X175/08 (Hampton Research HR2-814). You
just plunge your crystals in the liquid or wipe the crystal in the loop through
the liquid so that all excess water is removed.
Good luck !
Ulrike
...@utmb.edu
http://xray.utmb.edu
QQ: Research: If we got it right the first time it would just be called
search.
- A. Garcia
-Original Message-
From: Tim Gruene t...@shelx.uni-ac.gwdg.de
Reply-to: Tim Gruene t...@shelx.uni-ac.gwdg.de
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] cryo
bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] *On Behalf Of
*Ursula
Schulze-Gahmen
*Sent:* Monday, March 02, 2015 1:49 PM
*To:* CCP4BB@JISCMAIL.AC.UK
*Subject:* [ccp4bb] cryo protection for low salt crystallization at 4
degrees
I know there was jut recently a discussion about cryoconditions
I know there was jut recently a discussion about cryoconditions for
crystals, but I am still hoping for some new ideas for my crystals that
grow from HEPES buffer pH 7.3, 0.2 M NaCl by slowly lowering the
temperature from 20 to 4 degrees.
These crystals are easy to grow but extremely sensitive to
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Dear Ursula,
unless it is too long a list, maybe you could list the
cryo-protectants you have used so far? Did you try sugars (e.g.
glucose), or Na Malonate, or Butanediol (either 2,5 or 1,6, I don't
remember exactly)?
You could also increase the
What about 4deg data collection?
What about glutaraldehyde crosslinking?
JPK
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Ursula
Schulze-Gahmen
Sent: Monday, March 02, 2015 1:49 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] cryo protection for low salt crystallization
Hi All,
I have run into a very sensitive crystals system when it comes to cryo
protecting them. I have run through the usual suspects and trays are
going to be setup with a cryo protectant as part of crystallization
cocktail. The one problem that seems to be occurring is that the
you may have thought of this already, but you could try cryoprotection in the
drop itself.
i.e. slowly adding cryoprotectant to the reservoir, or replacing the reservoir
bit by bit with solution containing cryoprotectant, and then adding small
volumes to the side of the drop
- for example,
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Dear Len,
just to be on the safe side, my list of 'usual suspects' includes
- - glycerol/PEG400
- - LiCl et al at high concentration
- - Butanediol
- - sugars (glucose/ fructose)
- - oil
- - NaMalonate
- - MPD
...
you mention cracking upon
Len,
We have run into this problem from time to time, and it is very
frustrating. Here are some things to try, some of which you may have
done already:
Grow crystals in a small percentage of the cryoprotectant
(e.g., 5-10% glycerol). This often
[mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Leonard
Thomas
Sent: 26 October 2011 17:46
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] cryo protection
Hi All,
I have run into a very sensitive crystals system when it comes to cryo
protecting them. I have run through the usual suspects and trays are going
Hello Leonard,
one thing to test is whether transferring your crystals to a drop containing
simply well solution also causes cracking. If yes, then the possibility
exists that the absence of protein in solution is causing the trouble. In
that case, you can transfer the crystals to oil: you'll be
You can also try to crosslink before transferring to cryo.
From: Filip Van Petegem filip.vanpete...@gmail.com
To: CCP4BB@JISCMAIL.AC.UK CCP4BB@JISCMAIL.AC.UK
Sent: Wed Oct 26 13:19:16 2011
Subject: Re: [ccp4bb] cryo protection
Hello Leonard,
one thing to test
Hi Len;
I was having exactly the same problem with my crystals, but when we grow
the crystals in presence of increasing concentration of Glycerol and MPD
starting from 0.5 to 10%. The crystal doesn't appear after 3% of Glycerol
or MPD but the one which appear in 2.5 to 3 % were much resistant to
Another possibility (other than those already mentioned) is to try
freezing without a cryoprotectant, by fishing the crystals out onto a
mesh and removing all the mother liquor.
The following paper has some details:
Direct cryocooling of naked crystals: are cryoprotection agents
always
A good number of things to try. Just a little more info that was
asked for. The crystals are grown in Peg 3350 over a range of pH
values using Bis-Tris Propane. The are coming out of 2 different salt
conditions. My feeling is it is an osmolality problem though I also
observed cracking
Len,
May be you have already done this. I would closely check my
crystallization conditions and also check the pH of the cryo. In some
cases, during cryoprotection the pH of the original drop may drastically
different than the cryo solution. Also, sometime back, we were exploring
different
the crystals depending on the
concentration and the type of compounds in it.
Regards,
Mathews
-Original Message-
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Leonard
Thomas
Sent: Wednesday, October 26, 2011 11:57 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] cryo
One more thing you could try: high pressure cryo-cooling. Se any of a
number of paperas by Chae Un Kim; e.g.
http://www.ncbi.nlm.nih.gov/pubmed/17452791
Acta Crystallogr D Biol Crystallogr.
http://www.ncbi.nlm.nih.gov/pubmed/17452791# 2007 May;63(Pt 5):653-9.
Epub 2007 Apr 21.
On
Thomas
[lmtho...@ou.edu]
Sent: Wednesday, October 26, 2011 11:46 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] cryo protection
Hi All,
I have run into a very sensitive crystals system when it comes to cryo
protecting them. I have run through the usual suspects and trays are
...
Hey Len,
I had this problem, too. As you know, my favorite first try is always
fomblin (no need to mix anything). I had quite a bit success in stubborn
cases to inject about 4uL fomblin through the tape on top of the drop
and then looping crystals through the oil layer. You can wick the mother
I have always been a fan of oil, which has already been suggested. Have
you tried that?
Cross-linking has already been suggested, and these are some good protocols:
Lusty (1999) J. Appl. Crystallogr. 32, 106-112.
McWhirter, et al. (1999) PNAS USA 96, 8408-8413.
In the latter paper the
Dear Rongjin,
I would:
-prepare different cryosolutions, adding glycerol, replacing water
with glycerol, replacing ethanol with other more cryogenic alcohols:
if you have enough crystals, see how these behave when transferred to
these solutions, if they crack, decrease precipitant, if they
Dear All
I got crystals from 20% Ethanol with 0.1M Tris pH 8.5. This is my first
time to have crystals in Ethanol and want to get some suggestions of
cryo-protection from those who have done this before.
I am waiting for my time on home X-ray facility, and hope I can get
some suggestions before
Hi Rongjin,
I had crystals that grew in 9% ethanol as a precipitant. To cryoprotect
them, I first replaced the mother liquor with solution containing 10% MPD,
instead of ethanol, and then briefly dunked the crystals into ~30% MPD.
Hope this helps. Good luck.
Katya
On Mon, Sep 29, 2008 at 1:24
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