Re: [ccp4bb] suggestions for cryoprotectant

2018-10-20 Thread Joseph Brock 
Dear Friduous,


This is also a good paper to look at once you know your crystals diffract at 
room temperature:

https://www.ncbi.nlm.nih.gov/pubmed/10944336


Many of the salts in your condition will be cryo-protective if you increase 
their concentration to saturated levels.


Cheers,

Joseph.


Joseph S. Brock | PhD
Researcher
Drew Lab
Department of Biochemistry and Biophysics
Stockholm University

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Topics of the day:

  1. suggestions for cryoprotectant (4)
  2. RapiData at SSRL 2019



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Date:Fri, 19 Oct 2018 17:23:49 -0700
From:Christine Gee 
Subject: Re: suggestions for cryoprotectant

Dear Fridous

Have a look at https://www.ncbi.nlm.nih.gov/pubmed/14646118
[http://www.ncbi.nlm.nih.gov/coreutils/img/pubmed256blue.png]

Malonate: a versatile cryoprotectant and stabilizing solution for salt-grown 
macromolecular crystals. - PubMed - 
NCBI
www.ncbi.nlm.nih.gov
Acta Crystallogr D Biol Crystallogr. 2003 Dec;59(Pt 12):2356-8. Epub 2003 Nov 
27.


Malonate as a cryoprotectant

Or https://www.ncbi.nlm.nih.gov/pubmed/20606259
[http://www.ncbi.nlm.nih.gov/coreutils/img/pubmed256blue.png]

Cryoprotection properties of salts of organic acids: a case study for a 
tetragonal crystal of HEW lysozyme. - PubMed - 
NCBI
www.ncbi.nlm.nih.gov
Acta Crystallogr D Biol Crystallogr. 2010 Jul;66(Pt 7):789-96. doi: 
10.1107/S0907444910015416. Epub 2010 Jun 19.



Regards
Christine

On Fri, Oct 19, 2018 at 2:57 PM Firdous Tarique 
wrote:

> Dear members
>
> I have got beautiful crystal hits in SaltRx screens which are not
> diffracting to a good resoultion. All of them are salt based condition and
> I am not able to formulate a good cryoprotectant for these crystals. I also
> think that in my case the poor resolution is due to a poor cryoprotectant
> selection.
>
> The conditions are as follows:
>
> 1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0
> 2>0.5M KCN 100mM Tris pH8.5
> 3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0
> 4>4M Sodium Nitrate 100mM Tris pH8.5
> 5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6
>
> There are few more conditions but so far not able to see good diffraction
> with using lower peg and glycerol based cryoprotectants.
>
> Can anybody suggest me good cryos conditions for salt based
> crystallization conditions or anything good for SaltRx crystallization hits.
>
> Thanks
>
> Firdous
>
> --
>
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--

Date:Sat, 20 Oct 2018 02:21:41 +
From:"Smith, Clyde" 
Subject: RapiData at SSRL 2019

Hi everyone,

Applications are now open for the 2019 edition of RapiData at SSRL. RapiData is 
a practical course in macromolecular X-ray diffraction data collection, data 
processing and structure solution. The aim of the RapiData course is to educate 
and train young scientists in data collection and processing methods at 
synchrotron beamlines, using state-of-the-art software and instrumentation. The 
course will be held at the Stanford Synchrotron Radiation Lightsource (SSRL) 
located on the SLAC National Accelerator Laboratory campus in Menlo Park, CA, 
from May 5-10 2019. Course organizers for 2019 are Silvia Russi, Clyde Smith 
and Jeney Wierman.

The course will comprise hands-on experiments at the SSRL beamlines, software 
tutorials, and lectures on the following topics:
Specimen preparation, Data collection strategies, X-ray light sources, X-ray 
detectors, Data reduction, Structure solution by MAD, SAD and Molecular 
Replacement, Complementary methods (spectroscopy and small angle scattering)

Please visit http://smb.slac.stanford.edu/news/rapidata/rapidata-2019/ and use 
the links on the "registration" page to apply for the course. The application 
deadline is December 31 2018 Successful applicants will be notified early in 
January 2019 and invited to register and book accommodation at the SLAC 
Guesthouse at that time. A limited amount of travel support funding may be 
available.

[ccp4bb] Fw: Postdoctoral Fellow Position in Structural Biology

2018-10-20 Thread Vijay Parashar 
Postdoctoral Fellow Position
Parashar laboratory at University of Delaware College of Health Sciences, is 
currently seeking a Postdoctoral Fellow to perform structure-function studies 
of proteins regulating bacterial signaling.
 Qualifications:·  Ph.D. in Biochemistry, Molecular Biology, Structural 
Biology, or a related field·  Experience with cloning, protein expression, 
and protein purification methods·  A sound understanding of methods and 
approaches relating to macromolecular crystallography and/or single particle 
Cryo-EM preferred.  ·  Ability to plan and carry out research projects 
independently·  Ability to work on own initiative and take personal 
responsibility for delivery of work·  Excellent problem solving, 
interpersonal, communication and presentation skills·  A working knowledge 
of bacterial genetics and computational modeling of protein-ligand interactions 
preferred Responsibilities include:·  Performing experiments for cloning, 
protein expression, and purification of proteins for biochemical and structural 
studies; robotic screens for crystallization; optimizing crystallization 
conditions; and collecting and analyzing diffraction data.·  Performing 
model building and structure refinement.·  Maintaining laboratory equipment 
to meet unique needs of the research activity·  Assisting with preparation 
of grant proposals, presentations and publications·  Actively seeking new 
collaborative projects within the department and externally University of 
Delaware is well equipped with state-of-the-art equipment for molecular 
biology, protein purification, and crystallography, including crystallization 
robot, crystal imaging system, and X-ray generator. Please visit Parashar 
Laboratory website for more information. The laboratory plans to start using 
Cryo-EM for structural elucidation. For consideration, please send your CV, 
list of three references and other relevant information to paras...@udel.edu 
with subject line "postdoctoral application".
  



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Re: [ccp4bb] suggestions for cryoprotectant

2018-10-20 Thread Thomas, Leonard M. 
This is the first thing you need to do before going down the rabbit hole 
looking for a cryo protectant.  Mitegen make it much easier then in the 'old 
days' of capillaries.

Good luck,
Len

lmtho...@ou.edu

On Oct 20, 2018 2:02 AM, Andreas Förster  wrote:
Dear Firdous,

get some Mitegen sleeves and test your crystals at RT. Before you establish  
diffraction, there’s no need to agonize about cryo-conditions.

Good luck!


Andreas

On Fri, 19 Oct 2018 at 23:57, Firdous Tarique 
mailto:kahkashantari...@gmail.com>> wrote:
Dear members

I have got beautiful crystal hits in SaltRx screens which are not diffracting 
to a good resoultion. All of them are salt based condition and I am not able to 
formulate a good cryoprotectant for these crystals. I also think that in my 
case the poor resolution is due to a poor cryoprotectant selection.

The conditions are as follows:

1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0
2>0.5M KCN 100mM Tris pH8.5
3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0
4>4M Sodium Nitrate 100mM Tris pH8.5
5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6

There are few more conditions but so far not able to see good diffraction with 
using lower peg and glycerol based cryoprotectants.

Can anybody suggest me good cryos conditions for salt based crystallization 
conditions or anything good for SaltRx crystallization hits.

Thanks

Firdous



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Re: [ccp4bb] suggestions for cryoprotectant

2018-10-20 Thread mesters 

Hi,

have a look at the following interesting papers

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5466044/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2852300/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4252874/

Good luck

J.

Am 19.10.18 um 23:57 schrieb Firdous Tarique:

Dear members

I have got beautiful crystal hits in SaltRx screens which are not 
diffracting to a good resoultion. All of them are salt based condition 
and I am not able to formulate a good cryoprotectant for these 
crystals. I also think that in my case the poor resolution is due to a 
poor cryoprotectant selection.


The conditions are as follows:

1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0
2>0.5M KCN 100mM Tris pH8.5
3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0
4>4M Sodium Nitrate 100mM Tris pH8.5
5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6

There are few more conditions but so far not able to see good 
diffraction with using lower peg and glycerol based cryoprotectants.


Can anybody suggest me good cryos conditions for salt based 
crystallization conditions or anything good for SaltRx crystallization 
hits.


Thanks

Firdous



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Re: [ccp4bb] suggestions for cryoprotectant

2018-10-20 Thread Andreas Förster 
Dear Firdous,

get some Mitegen sleeves and test your crystals at RT. Before you establish
 diffraction, there’s no need to agonize about cryo-conditions.

Good luck!


Andreas

On Fri, 19 Oct 2018 at 23:57, Firdous Tarique 
wrote:

> Dear members
>
> I have got beautiful crystal hits in SaltRx screens which are not
> diffracting to a good resoultion. All of them are salt based condition and
> I am not able to formulate a good cryoprotectant for these crystals. I also
> think that in my case the poor resolution is due to a poor cryoprotectant
> selection.
>
> The conditions are as follows:
>
> 1> 4M Ammonium Acetate 100mM Bis Tris Propane pH 7.0
> 2>0.5M KCN 100mM Tris pH8.5
> 3>1.5M LiSo4 100mM Bris Tris Propane pH 7.0
> 4>4M Sodium Nitrate 100mM Tris pH8.5
> 5>1.5M Sodium Nitrate 100mM Sodium acetate pH 4.6
>
> There are few more conditions but so far not able to see good diffraction
> with using lower peg and glycerol based cryoprotectants.
>
> Can anybody suggest me good cryos conditions for salt based
> crystallization conditions or anything good for SaltRx crystallization hits.
>
> Thanks
>
> Firdous
>
> --
>
> To unsubscribe from the CCP4BB list, click the following link:
> https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB=1
>



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