Hi all
Just wanted to thank you all for all the suggestions and tell you of all
the things i tried what finally worked, if anyones interested.
Among the several things mentioned here, the crystals diffracted the best
(~ 2.9 Å at the synchrotron) when grown in a solution containing either 5%
ethyl
Dear CCP4
Thank you for your suggestions
Best Regards
Rana
Hi Rana,
you are almost there. For the first one I'd add (if at all) 5% PEG200
and for the second I'd mix in 10% PEG200.
Only very big crystals might need even more cryoprotectant.
Best, Matthias
-
Dr. Matthias Zebisch
Division of Structural Biology,
W
Dear CCP4
Does anyone know a good cryoprotection condition for these two conditions I
would be grateful
The first condition:
10% PEG 2, 20%PEG MME 550. 0.03M of NPS ( NPS is sodium nitrate, disodium
hydrogen phosphate, and ammonium sulfate), 0.1MMES/Imadizole pH 6.5
The second condition:
20%
cryo-protectants you have already tried. For the ones that work, repeat
> with higher amounts. Very often crystals are much happier when you don't
> soak/tinker with them and freeze them straight out of the drop.
>
> Good luck.
>
> Mark
>
>
>
> -Original
araju
To: CCP4BB
Sent: Wed, Jan 22, 2014 10:38 am
Subject: [ccp4bb] cryoprotection ideas for salt based condition
Hello folks,
I have crystals for a protein which form in 0.1 M MES pH 6-6.75, 10% dioxane
and 1.6-2 M Ammonium sulphate. Based on little experience I have finding the
Mahesh,
Try 25-30% glucose. You can gradually add well solution + 37.5-40%
glucose directly to the drop if your crystals are sensitive to changes
in osmotic pressure. If your drop is evaporating too quickly, try
working in the cold room or under oil to slow down evaporation. You can
find our
Hello folks,
I have crystals for a protein which form in 0.1 M MES pH 6-6.75, 10%
dioxane and 1.6-2 M Ammonium sulphate. Based on little experience I have
finding the right cryoprotection for salt based conditions; I have tried
glycerol (15-25%), Ethylene glycol (20-25%), DMSO (15-20%) and increas
@JISCMAIL.AC.UK] Im Auftrag von Enrico
Stura
Gesendet: Dienstag, 27. August 2013 11:58
An: CCP4BB@JISCMAIL.AC.UK
Betreff: Re: [ccp4bb] AW: [ccp4bb] cryoprotection
Herman,
The trick you suggest is not as valid as you may think. The ice rings can
originate from the crystal itself.
If you crystallize in
itag, 23. August 2013 19:52
An: CCP4BB@JISCMAIL.AC.UK
Betreff: [ccp4bb] cryoprotection
Hello
Can anyone suggest a cryoprotectant for the following crystallization
condition
0.2-0.4M sodium formate
~20% PEG 3350
0-25 mM Nickel
0-100 mM Malonate
Thank you
with regards
uday
--
Enrico A. S
Nachricht-
Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] Im Auftrag von Uday
Kumar
Gesendet: Freitag, 23. August 2013 19:52
An: CCP4BB@JISCMAIL.AC.UK
Betreff: [ccp4bb] cryoprotection
Hello
Can anyone suggest a cryoprotectant for the following crystallization condition
0.2-0.4M sodium
Dear uday,
I have grown crystals in 0.2M sodium formate and 20% peg3350, i tried cryo
solution cotaining 0.2M sodium formate and peg1050 40%, it worked well for
me and diffracted to 2.3A at home source.
0.2-0.4M sodium formate
40% PEG 1050
0-25 mM Nickel
0-100 mM Malonate
On Fri, Aug 23, 2013
I've had good success transferring crystals grown in
200 mM NaOAc pH 5.5 / 10% PEG
Into a solution for room temperature diffraction (in ~6 steps, peg first, then
buffers) :
100 mM NaOAc / 100 mM Malonate / 30% PEG
And from there into a cryo solution in 4-5 steps) that was:
100 mM NaOAc / 100
increase your PEG3350 >27% and keep the other components at their current
concentration. You can also add glycerol or ethylene glycol into the mix.
If you have multiple crystals then try various variants.
Jürgen
On Aug 23, 2013, at 1:52 PM, Uday Kumar wrote:
Hello
Can anyone suggest a cryopro
Hello
Can anyone suggest a cryoprotectant for the following crystallization condition
0.2-0.4M sodium formate
~20% PEG 3350
0-25 mM Nickel
0-100 mM Malonate
Thank you
with regards
uday
I believe this is the article that you are looking for:
http://www.xtals.org/crystal_cryo.pdf
Cheers,
Sean
I found within the last few years a nice flowchart that someone (I think it
was Artem Evdokimov) had posted somewhere for a protocol for dealing with
crystals that you couldn't just quick dip, and now I can't find it - maybe
he can send it to me offbb??? Thanks
Laurie Betts
UNC CHapel Hill
Hi Brenda,
Maybe you want to have a look at the cryoprotectant database for
protein crystals.
http://idb.exst.jaxa.jp/db_data/protein/search-e.php
There you can search for your precipitant and get a list of
compatible freezing conditions that worked before including the
reference. It is
: +32-
(0)9-264.51.24 ; mobile: +32-(0)472-92.85.19 Email: savvas.savvi...@ugent.be
http://www.lprobe.ugent.be/xray.html
-Original Message-
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
Schulman, Brenda
Sent: Wednesday, August 19, 2009 6:19 PM
To: CC
bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
Schulman, Brenda
Sent: Wednesday, August 19, 2009 6:19 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Cryoprotection in > 3M ammonium sulfate
Hello!
I would be grateful for suggestions on cryoprotectants for crystals growing
in > 3M a
bulletin board [ccp...@jiscmail.ac.uk] On Behalf Of Savvas Savvides
[savvas.savvi...@ugent.be]
Sent: Wednesday, August 19, 2009 11:47 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Cryoprotection in > 3M ammonium sulfate
Hi Brenda
Try > 3M ammonium sulfate itself! We have tried that with
ulman, Brenda
Sent: Wednesday, August 19, 2009 12:19 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Cryoprotection in > 3M ammonium sulfate
Hello!
I would be grateful for suggestions on cryoprotectants for crystals
growing
in > 3M ammonium sulfate.
Thanks!
Brenda
Email Disclaimer: www
-Original Message-
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of
Schulman, Brenda
Sent: Wednesday, August 19, 2009 6:19 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Cryoprotection in > 3M ammonium sulfate
Hello!
I would be grateful for suggestions on cryoprot
Dear Brenda
Check out malonate
http://www.ncbi.nlm.nih.gov/pubmed/14646118?
ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubm
ed_DefaultReportPanel.Pubmed_RVDocSum
Malonate: a versatile cryoprotectant and stabilizing solution for
salt-grown macromolecular crystals.
H
Soluble possibilities include glucose, ethylene glycol, DMSO, glycerol
(15-30%). You may not need much cryoprotectant at all at these salt
concentrations. PEGs are not very soluble in high ammonium sulfate
solutions. (I think PEG-400 can go up to only 4% or so at 2 M ammonium
sulfate.)
Cheers
Hi Brenda,
You can try sugars like glucose, trehalose and sucrose for high AS contents.
It has been succesfully used in really hard cases such at protein RNA
crystals grown in AS. see Acta Cryst (2002) D58 1664-1669 Garber et al.
HTH
Pascal Egea
Hello!
I would be grateful for suggestions on cryoprotectants for crystals growing
in > 3M ammonium sulfate.
Thanks!
Brenda
Email Disclaimer: www.stjude.org/emaildisclaimer
27 matches
Mail list logo
