Along with all the excellent suggestions so far you can also try no
cryoprotectant at all: If you harvest your crystal on a mesh loop and
remove all the mother liquor the crystal lattice itself will act as a
cryoprotectant as long as the solvent channels are smaller than 40A in
diameter (they
Hi Faisal,
Did you try to simply raise the Sokalan CP7 percentage? Additives like
Glycerol can increase the solubility of proteins, in that case you
have to counteract by increasing also the precipitant concentration.
If your crystals crack, a likely reason is osmotic shock. Particularly
Hello everyone
Can anybody suggest me a cryo condition for a crystal obtained in
MIDAS screen of Molecular Dimension:
G1 0.1MTris8.0G10.1Mpotassium chloride25% v/vSOKALAN®CP7 0.1MHEPES7.0
G20.3Mammonium formate20% v/vSOKALAN® CP 5 0.1MHEPES7.0
Crystals are in beautiful cuboid shaped but all
like you describe.
GL Bert
From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Faisal Tarique
[faisaltari...@gmail.com]
Sent: Monday, May 04, 2015 7:43 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] cryo condition
Hello everyone
Can anybody
of Roger
Rowlett rrowl...@colgate.edu
Sent: Tuesday, 5 May 2015 7:22 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] cryo condition
We rarely use glycerol anymore, because it seems to fail so often for
many of our current proteins. Try glucose, 25-30%. This is most
conveniently done
What about nature's favourite cryoprotectant, trehalose?
From: CCP4 bulletin board CCP4BB@JISCMAIL.AC.UK on behalf of Roger Rowlett
rrowl...@colgate.edu
Sent: Tuesday, 5 May 2015 7:22 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] cryo condition
We
We rarely use glycerol anymore, because it seems to fail so often for
many of our current proteins. Try glucose, 25-30%. This is most
conveniently done by weighing 125-150 mg of glucose in a microcentrifuge
tube, then addding well solution to the 0.5 mL mark and mixing until
completely
If in doubt, try dragging the crystal through a 1:1 mix of Paratone-N and
Mineral oil until most or all the mother liquor from surrounding the
crystal has been removed.
There are more tips here:
http://hamptonresearch.com/tip_detail.aspx?id=99
Good luck!
D
On Mon, 4 May 2015 19:45 Faisal
Thank you everybody for their nice suggestions..
On Thu, May 23, 2013 at 7:39 PM, Matthew BOWLER mbow...@embl.fr wrote:
I keep sending mails by accident today - apologies for the spam. The last
sentence of my should read:
This could of course be due to too high a concentration of mother
Dear all
Can anybody tell me the appropriate cryo condition for the crystals
obtained in 2M Ammonium sulphate and tris pH8.5..I tried to add 10%
glycerol to it but still the ice ring is forming..
Thanx in advance
--
Regards
Faisal
School of Life Sciences
JNU
80% saturated Li2SO4
On Thu, 23 May 2013 11:42:09 +0200, Faisal Tarique
faisaltari...@gmail.com wrote:
Dear all
Can anybody tell me the appropriate cryo condition for the crystals
obtained in 2M Ammonium sulphate and tris pH8.5..I tried to add 10%
glycerol to it but still the ice ring is
From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Faisal Tarique [faisaltari...@gmail.com]
Sent: Thursday, May 23, 2013 12:42 PM
To:
CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] cryo condition
Dear all
Can anybody tell me the appropriate cryo condition for the crystals obtained
30% glycerol or 25% glucose should be sufficient for 1-2 M ammonium sulfate.
Roger Rowlett
On May 23, 2013 5:42 AM, Faisal Tarique faisaltari...@gmail.com wrote:
Dear all
Can anybody tell me the appropriate cryo condition for the crystals
obtained in 2M Ammonium sulphate and tris pH8.5..I
From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Roger Rowlett
[rrowl...@colgate.edu]
Sent: 23 May 2013 21:21
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] cryo condition
30% glycerol or 25% glucose should be sufficient for 1-2 M ammonium sulfate.
Roger Rowlett
On May 23, 2013 5:42
Hi Faisal,
3 - 3.5M Ammonium sulphate with your buffer should work too.
Take a small loop.
Jan
--
Jan Abendroth
Emerald BioStructures
Seattle / Bainbridge Island WA, USA
home: Jan.Abendroth_at_gmail.com
work: JAbendroth_at_embios.com
http://www.emeraldbiostructures.com
On May 23, 2013, at 2:42
Hi Faisal,
On 2013-05-23 11:42, Faisal Tarique wrote:
Dear all
Can anybody tell me the appropriate cryo condition for the crystals
obtained in 2M Ammonium sulphate and tris pH8.5..I tried to add 10%
glycerol to it but still the ice ring is forming..
Thanx in advance--
Regards
Faisal
School
Hi Faisal,
if your solvent channels are smaller than 40A in the largest dimension
(most are) you can use a mesh loop to pick up the crystal and then wick
away all of the mother liquor. You can then flash cool your crystal
without having to transfer the crystal to another solution. Good luck,
: 42.2 GHz
Heisenberg was probably here!
-Original Message-
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Thibaut
Crepin
Sent: Thursday, May 23, 2013 6:24 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] cryo condition
Dear Faisal,
this paper can be really
L-proline works well with ammonium sulfate:
http://www.ncbi.nlm.nih.gov/pubmed/22868767
Sent from Jack's iPad
On May 23, 2013, at 4:42 AM, Faisal Tarique faisaltari...@gmail.com wrote:
Dear all
Can anybody tell me the appropriate cryo condition for the crystals obtained
in 2M Ammonium
Matt,
with this technique, how do you prevent crystal from drying up (other
than doing it fast)? I know Thorne's group does this trick under oil.
If you take no extra precautions, do you have an estimate of how often
diffraction is destroyed by this?
On the other hand, it's quite possible
Hi Ed,
good question. I have found that you have a good 30 seconds to remove
the surrounding liquid - so while you have to do it fast you have enough
time that it doesn't need a robot and even a malcoordinate such as
myself can do it. I'm afraid that I have no estimate for how often
I keep sending mails by accident today - apologies for the spam. The
last sentence of my should read:
This could of course be due to too high a concentration of mother
liquor but quite often it occurs at relative humidity values where the
concentration of the mother liquor components will
22 matches
Mail list logo
