Re: [ccp4bb] need some suggestions for crystallization

[Jacob Keller]

Does that CA have a metal center (I think they all do)? If so, doesn't the
citrate compete for the metal?

JPK

On Mon, Feb 4, 2013 at 2:25 PM, Roger Rowlett  wrote:

> Human carbonic anhdyrase II can be easily crystallized from 1.3 M sodium
> citrate/0.1 M TrisCl pH 8.5 at 10 mg/mL protein concentration. Crystals are
> P21 and easily diffract to beyond 2.0 A on a home source. We cryopreserve
> in ML + 30% glucose. Sulfonamide ligands are easy to soak into the crystals
> in a few minutes during cryopreservation, and provide teaching
> opportunities for protein-ligand model building and refinement.
>
> Cheers,
>
> __**_
> Roger Rowlett
> Gordon & Dorothy Kline Professor
> Department of Chemistry
> Colgate University
> 13 Oak Drive
> Hamilton, NY 13346
>
> tel: (315)-228-7245
> ofc: (315)-228-7395
> fax: (315)-228-7935
> email: rrowl...@colgate.edu
>
>
> On 02/04/2013 12:31 PM, Jim Pflugrath wrote:
>
>> A number of protein crystal recipes are available on the Rigaku web site:
>> http://www.rigaku.com/**products/protein/recipes
>>
>> Lysozyme is nice because it is so cheap, grows quickly, cryoprotectants
>> in a straightforward way, and the unit cells are not large nor are the
>> crystals fragile.  One can get triclinic, monoclinic, orthorhombic, and
>> tetragonal crystals relatively quickly.
>>
>> With hen egg white lysozyme if you are not up for sulfur-SAD phasing,
>> then co-crystallizing or quick-soaking in iodide (say 100 mM) gives a very
>> nice anomalous signal at just about any wavelength.  That is, no need to
>> worry about going to a so-called edge.
>>
>> While there are many other easy-to-go crystals, I have found that none of
>> them combine all the properties of hen egg white lysozyme has a good
>> teaching tool.
>>
>> Jim
>>
>> ==
>> Can you all suggest some protein crystallization conditions (along with
>> cryo conditions) for some commercially available proteins?  I'm looking
>> to get 6-8 different ones (and we'll just take them and see how it
>> goes).  I wouldn't mind knowing unit cell parameters as well (just a
>> citation works, I can have them figure it out).  I have about 7 weeks to
>> get everything grown and frozen and ready to go.
>>
>> Any help would be greatly appreciated.  It always amazes me how helpful
>> this group is.  Thank you very much.
>>
>> Dave
>>
>


-- 
***
Jacob Pearson Keller, PhD
Postdoctoral Associate
HHMI Janelia Farms Research Campus
email: j-kell...@northwestern.edu
***

Re: [ccp4bb] need some suggestions for crystallization

[Michael Thompson]

Hi Dave,

ProteinaseK is also a good one. Crystallizes rapidly, big crystals, and 
relatively high resolution data (1.0-1.5A) usually. You can also buy the 
lyophilized powder from sigma and prepare the sample directly from the 
commercial material. We use proK for a course here at UCLA, so if you are 
interested I can send you more details about the protocol.

Mike



- Original Message -
From: "David Roberts" 
To: CCP4BB@JISCMAIL.AC.UK
Sent: Monday, February 4, 2013 8:03:09 AM GMT -08:00 US/Canada Pacific
Subject: [ccp4bb] need some suggestions for crystallization

So, I know I say this every time I post on this board, but here it goes 
again.

I'm at an undergrad only school, and every 2 years I teach a class in 
protein crystallography.  This year I'm being super ambitious, and I'm 
going to take a class of 16 to the synchrotron for data collection.  
It's just an 8 hour thing, to show them the entire process.  I'm hoping 
that we can collect 5-6 good data sets while there.

I would like them to grow their own crystals, and go collect data. Then 
we'd come back and actually do a molecular replacement (pretty 
easy/standard really).  Just to get a feel for how it works.

The protein I do research on is not one that I would push on this, as 
the crystals are hard to grow, they are very soft, and the data just 
isn't the best (resolution issues).  I do have a few that will work on 
my proteins, but I was thinking of having others in the class grow up 
classic proteins for data collection.  Obviously lysozyme is one, but I 
was wondering what other standard bulletproof conditions are out there.

Can you all suggest some protein crystallization conditions (along with 
cryo conditions) for some commercially available proteins?  I'm looking 
to get 6-8 different ones (and we'll just take them and see how it 
goes).  I wouldn't mind knowing unit cell parameters as well (just a 
citation works, I can have them figure it out).  I have about 7 weeks to 
get everything grown and frozen and ready to go.

Any help would be greatly appreciated.  It always amazes me how helpful 
this group is.  Thank you very much.

Dave

-- 
Michael C. Thompson

Graduate Student

Biochemistry & Molecular Biology Division

Department of Chemistry & Biochemistry

University of California, Los Angeles

mi...@chem.ucla.edu

Re: [ccp4bb] need some suggestions for crystallization

[Roger Rowlett]

Human carbonic anhdyrase II can be easily crystallized from 1.3 M sodium 
citrate/0.1 M TrisCl pH 8.5 at 10 mg/mL protein concentration. Crystals 
are P21 and easily diffract to beyond 2.0 A on a home source. We 
cryopreserve in ML + 30% glucose. Sulfonamide ligands are easy to soak 
into the crystals in a few minutes during cryopreservation, and provide 
teaching opportunities for protein-ligand model building and refinement.


Cheers,

___
Roger Rowlett
Gordon & Dorothy Kline Professor
Department of Chemistry
Colgate University
13 Oak Drive
Hamilton, NY 13346

tel: (315)-228-7245
ofc: (315)-228-7395
fax: (315)-228-7935
email: rrowl...@colgate.edu

On 02/04/2013 12:31 PM, Jim Pflugrath wrote:

A number of protein crystal recipes are available on the Rigaku web site:
http://www.rigaku.com/products/protein/recipes

Lysozyme is nice because it is so cheap, grows quickly, cryoprotectants in a 
straightforward way, and the unit cells are not large nor are the crystals 
fragile.  One can get triclinic, monoclinic, orthorhombic, and tetragonal 
crystals relatively quickly.

With hen egg white lysozyme if you are not up for sulfur-SAD phasing, then 
co-crystallizing or quick-soaking in iodide (say 100 mM) gives a very nice 
anomalous signal at just about any wavelength.  That is, no need to worry about 
going to a so-called edge.

While there are many other easy-to-go crystals, I have found that none of them 
combine all the properties of hen egg white lysozyme has a good teaching tool.

Jim

==
Can you all suggest some protein crystallization conditions (along with
cryo conditions) for some commercially available proteins?  I'm looking
to get 6-8 different ones (and we'll just take them and see how it
goes).  I wouldn't mind knowing unit cell parameters as well (just a
citation works, I can have them figure it out).  I have about 7 weeks to
get everything grown and frozen and ready to go.

Any help would be greatly appreciated.  It always amazes me how helpful
this group is.  Thank you very much.

Dave

Re: [ccp4bb] need some suggestions for crystallization

[Jim Pflugrath]

A number of protein crystal recipes are available on the Rigaku web site:
http://www.rigaku.com/products/protein/recipes

Lysozyme is nice because it is so cheap, grows quickly, cryoprotectants in a 
straightforward way, and the unit cells are not large nor are the crystals 
fragile.  One can get triclinic, monoclinic, orthorhombic, and tetragonal 
crystals relatively quickly.

With hen egg white lysozyme if you are not up for sulfur-SAD phasing, then 
co-crystallizing or quick-soaking in iodide (say 100 mM) gives a very nice 
anomalous signal at just about any wavelength.  That is, no need to worry about 
going to a so-called edge.

While there are many other easy-to-go crystals, I have found that none of them 
combine all the properties of hen egg white lysozyme has a good teaching tool.

Jim

==
Can you all suggest some protein crystallization conditions (along with
cryo conditions) for some commercially available proteins?  I'm looking
to get 6-8 different ones (and we'll just take them and see how it
goes).  I wouldn't mind knowing unit cell parameters as well (just a
citation works, I can have them figure it out).  I have about 7 weeks to
get everything grown and frozen and ready to go.

Any help would be greatly appreciated.  It always amazes me how helpful
this group is.  Thank you very much.

Dave

Re: [ccp4bb] need some suggestions for crystallization

[vellieux]

I second that. Gently swirling the bottle (very important) before 
pipetting a few microliters (say 100 microliters or whatever).


Dialyse overnight vs 10 mM HEPES 2 mM MgCl2 ph7, then get the protein 
concentration to 10 mg/ml. Set up the drops (sitting drops) versus the 
same buffer with 30% (v/v) MPD. Direct cryo conditions. Depending on the 
material (plastic) used for the sitting drop trays you may find it 
difficult to get crystals out - you'll break up a few - but then no 
messing up to try to find cryo-conditions.


Fred.

On 04/02/13 17:27, Ganesh Natrajan wrote:

Hi David,

You could try the Glucose Isomerase supplied by Hampton. It 
crystallizes under a number of conditions, details of which you can 
find in their manual.



http://hamptonresearch.com/product_detail.aspx?cid=28&sid=56&pid=56

Ganesh


--
Fred. Vellieux (B.Sc., Ph.D., hdr)
IBS / ELMA
41 rue Jules Horowitz
F-38027 Grenoble Cedex 01
Tel: +33 438789605
Fax: +33 438785494

Re: [ccp4bb] need some suggestions for crystallization

[Radisky, Evette S., Ph.D.]

Bovine trypsin works well.  You can buy it pretty cheap from Sigma and it 
crystallizes without further purification, within a week.  Crystals diffract to 
1.1-1.3 A and are quite robust to handling and soaking.  Conditions that I used 
are described in this ref:
http://www.pnas.org/content/103/18/6835.long

I would avoid commercial preps of porcine elastase; I messed around with that 
one around 10 years ago using many reported crystallization conditions and 
suppliers but never got crystals.

Evette S. Radisky, Ph.D.
Assistant Professor
Mayo Clinic Cancer Center
Griffin Cancer Research Building, Rm 310
4500 San Pablo Road
Jacksonville, FL 32224
(904) 953-6372

-Original Message-
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of David 
Roberts
Sent: Monday, February 04, 2013 11:03 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] need some suggestions for crystallization

So, I know I say this every time I post on this board, but here it goes again.

I'm at an undergrad only school, and every 2 years I teach a class in protein 
crystallography.  This year I'm being super ambitious, and I'm going to take a 
class of 16 to the synchrotron for data collection.  
It's just an 8 hour thing, to show them the entire process.  I'm hoping that we 
can collect 5-6 good data sets while there.

I would like them to grow their own crystals, and go collect data. Then we'd 
come back and actually do a molecular replacement (pretty easy/standard 
really).  Just to get a feel for how it works.

The protein I do research on is not one that I would push on this, as the 
crystals are hard to grow, they are very soft, and the data just isn't the best 
(resolution issues).  I do have a few that will work on my proteins, but I was 
thinking of having others in the class grow up classic proteins for data 
collection.  Obviously lysozyme is one, but I was wondering what other standard 
bulletproof conditions are out there.

Can you all suggest some protein crystallization conditions (along with cryo 
conditions) for some commercially available proteins?  I'm looking to get 6-8 
different ones (and we'll just take them and see how it goes).  I wouldn't mind 
knowing unit cell parameters as well (just a citation works, I can have them 
figure it out).  I have about 7 weeks to get everything grown and frozen and 
ready to go.

Any help would be greatly appreciated.  It always amazes me how helpful this 
group is.  Thank you very much.

Dave

Re: [ccp4bb] need some suggestions for crystallization

[Tim Gruene]

-BEGIN PGP SIGNED MESSAGE-
Hash: SHA1

Dear Dave,

many methods articles mention a small set of commonly used proteins. E.g.
Mueller et al, "Optimal fine phi-slicing for single-photon-counting
pixel detectors", Acta Cryst D68, p42-56 list Insulin, Lysozyme,
Thaumatin, and Thermolysin;

Nanao et al, "Improving radiation-damage substructures for RIP", Acta
D61, 1227-1237, list Elastase, Insulin, Lysozyme, Ribonuclease A,
Thaumatin, and Trypsin.
Elastase, though, decided not to crystallise any longer about ten'ish
years ago.

Attached is a different condition for Lysozyme which I received from
Prof. Susana Andrade and which I usually use for teaching. - the
crystals grow during lunch time. They can be picked directly and you
can show the cryo-protecting effect of Ethylene glycol with increasing
concentration.

Best,
Tim

On 02/04/2013 05:03 PM, David Roberts wrote:
> So, I know I say this every time I post on this board, but here it 
> goes again.
> 
> I'm at an undergrad only school, and every 2 years I teach a class 
> in protein crystallography.  This year I'm being super ambitious, 
> and I'm going to take a class of 16 to the synchrotron for data 
> collection. It's just an 8 hour thing, to show them the entire 
> process.  I'm hoping that we can collect 5-6 good data sets while 
> there.
> 
> I would like them to grow their own crystals, and go collect data. 
> Then we'd come back and actually do a molecular replacement (pretty
> easy/standard really).  Just to get a feel for how it works.
> 
> The protein I do research on is not one that I would push on this, 
> as the crystals are hard to grow, they are very soft, and the data 
> just isn't the best (resolution issues).  I do have a few that
> will work on my proteins, but I was thinking of having others in
> the class grow up classic proteins for data collection.  Obviously 
> lysozyme is one, but I was wondering what other standard 
> bulletproof conditions are out there.
> 
> Can you all suggest some protein crystallization conditions (along 
> with cryo conditions) for some commercially available proteins? I'm
> looking to get 6-8 different ones (and we'll just take them and see
> how it goes).  I wouldn't mind knowing unit cell parameters as well
> (just a citation works, I can have them figure it out).  I have
> about 7 weeks to get everything grown and frozen and ready to go.
> 
> Any help would be greatly appreciated.  It always amazes me how 
> helpful this group is.  Thank you very much.
> 
> Dave
> 

- -- 
- --
Dr Tim Gruene
Institut fuer anorganische Chemie
Tammannstr. 4
D-37077 Goettingen

GPG Key ID = A46BEE1A

-BEGIN PGP SIGNATURE-
Version: GnuPG v1.4.12 (GNU/Linux)
Comment: Using GnuPG with Mozilla - http://enigmail.mozdev.org/

iD8DBQFRD+M2UxlJ7aRr7hoRAnDhAJ0bE6xl0XDxV6qoKslThPyyVd3CwACgpjnA
sIjr3296ejuwby0ZyXLw5po=
=JgwT
-END PGP SIGNATURE-


xtalization_lysozyme.ods
Description: application/vnd.oasis.opendocument.spreadsheet

Re: [ccp4bb] need some suggestions for crystallization

[Raji Edayathumangalam]

My suggestions would be to look up citations for thaumatin and glucose
isomerase. If I remember correctly, both of them form well diffracting
crystals within a short period of time. I think you can also buy the
purified protein from a vendor. Perhaps you could also try the good old
lysozyme.

Cheers,
Raji



On Mon, Feb 4, 2013 at 11:03 AM, David Roberts  wrote:

> So, I know I say this every time I post on this board, but here it goes
> again.
>
> I'm at an undergrad only school, and every 2 years I teach a class in
> protein crystallography.  This year I'm being super ambitious, and I'm
> going to take a class of 16 to the synchrotron for data collection.  It's
> just an 8 hour thing, to show them the entire process.  I'm hoping that we
> can collect 5-6 good data sets while there.
>
> I would like them to grow their own crystals, and go collect data. Then
> we'd come back and actually do a molecular replacement (pretty
> easy/standard really).  Just to get a feel for how it works.
>
> The protein I do research on is not one that I would push on this, as the
> crystals are hard to grow, they are very soft, and the data just isn't the
> best (resolution issues).  I do have a few that will work on my proteins,
> but I was thinking of having others in the class grow up classic proteins
> for data collection.  Obviously lysozyme is one, but I was wondering what
> other standard bulletproof conditions are out there.
>
> Can you all suggest some protein crystallization conditions (along with
> cryo conditions) for some commercially available proteins?  I'm looking to
> get 6-8 different ones (and we'll just take them and see how it goes).  I
> wouldn't mind knowing unit cell parameters as well (just a citation works,
> I can have them figure it out).  I have about 7 weeks to get everything
> grown and frozen and ready to go.
>
> Any help would be greatly appreciated.  It always amazes me how helpful
> this group is.  Thank you very much.
>
> Dave
>



-- 
Raji Edayathumangalam
Instructor in Neurology, Harvard Medical School
Research Associate, Brigham and Women's Hospital
Visiting Research Scholar, Brandeis University

Re: [ccp4bb] need some suggestions for crystallization

[Dr. Anthony Addlagatta]

***
This message has been scanned by the InterScan for CSC SSM by IICT security 
policy and found to be free of known security risks.
***


Dave,

You can try any or all of these proteins commercially available and all 
conditions for
crystallization and freezing established.

LysozymeFerritinGlucose isomerase   Myoglobin
Proteinase KThaumatin   Trypsin

You can follow this link. We used some of the conditions mentioned in their 
successfully.

http://www.rigaku.com/products/protein/recipes

Disclaimer: I do not have any commercial interest with Rigaku.

Anthony

-
Dr. Anthony Addlagatta
Center for Chemical Biology 
Indian Institute of Chemical Technology [IICT]
Tarnaka, Hyderabad
AP-500 607, INDIA
Tel:91-40-27191812
Web: https://sites.google.com/site/chembioliict/home/dr-anthony-addlagatta-1

-- Original Message ---
From: Harry Powell 
To: CCP4BB@JISCMAIL.AC.UK
Sent: Mon, 4 Feb 2013 16:10:54 +
Subject: Re: [ccp4bb] need some suggestions for crystallization

> ***
> This message has been scanned by the InterScan for CSC SSM at IICT and found 
> to be
free of known security risks.
> ***
> 
> Hi David
> 
> try going back to the one that started it all,* myoglobin, a recipe  
> is at
> 
>   http://www.rigaku.com/products/protein/recipes
> 
> (* feel free to argue about this)
> 
> On 4 Feb 2013, at Mon4 Feb 16:03, David Roberts wrote:
> 
> > So, I know I say this every time I post on this board, but here it  
> > goes again.
> >
> > I'm at an undergrad only school, and every 2 years I teach a class  
> > in protein crystallography.  This year I'm being super ambitious,  
> > and I'm going to take a class of 16 to the synchrotron for data  
> > collection.  It's just an 8 hour thing, to show them the entire  
> > process.  I'm hoping that we can collect 5-6 good data sets while  
> > there.
> >
> > I would like them to grow their own crystals, and go collect data.  
> > Then we'd come back and actually do a molecular replacement (pretty  
> > easy/standard really).  Just to get a feel for how it works.
> >
> > The protein I do research on is not one that I would push on this,  
> > as the crystals are hard to grow, they are very soft, and the data  
> > just isn't the best (resolution issues).  I do have a few that will  
> > work on my proteins, but I was thinking of having others in the  
> > class grow up classic proteins for data collection.  Obviously  
> > lysozyme is one, but I was wondering what other standard  
> > bulletproof conditions are out there.
> >
> > Can you all suggest some protein crystallization conditions (along  
> > with cryo conditions) for some commercially available proteins?   
> > I'm looking to get 6-8 different ones (and we'll just take them and  
> > see how it goes).  I wouldn't mind knowing unit cell parameters as  
> > well (just a citation works, I can have them figure it out).  I  
> > have about 7 weeks to get everything grown and frozen and ready to go.
> >
> > Any help would be greatly appreciated.  It always amazes me how  
> > helpful this group is.  Thank you very much.
> >
> > Dave
> 
> Harry
> --
> Dr Harry Powell, MRC Laboratory of Molecular Biology, MRC Centre,  
> Hills Road, Cambridge, CB2 0QH
> Chairman of European Crystallographic Association SIG9  
> (Crystallographic Computing)
--- End of Original Message ---

This Mail Scanned by ClamAV and Spammassassin

Re: [ccp4bb] need some suggestions for crystallization

[Ganesh Natrajan]

Hi David,

You could try the Glucose Isomerase supplied by Hampton. It crystallizes 
under a number of conditions, details of which you can find in their manual.



http://hamptonresearch.com/product_detail.aspx?cid=28&sid=56&pid=56

Ganesh



Le 04/02/13 17:03, David Roberts a écrit :
So, I know I say this every time I post on this board, but here it 
goes again.


I'm at an undergrad only school, and every 2 years I teach a class in 
protein crystallography.  This year I'm being super ambitious, and I'm 
going to take a class of 16 to the synchrotron for data collection.  
It's just an 8 hour thing, to show them the entire process.  I'm 
hoping that we can collect 5-6 good data sets while there.


I would like them to grow their own crystals, and go collect data. 
Then we'd come back and actually do a molecular replacement (pretty 
easy/standard really).  Just to get a feel for how it works.


The protein I do research on is not one that I would push on this, as 
the crystals are hard to grow, they are very soft, and the data just 
isn't the best (resolution issues).  I do have a few that will work on 
my proteins, but I was thinking of having others in the class grow up 
classic proteins for data collection.  Obviously lysozyme is one, but 
I was wondering what other standard bulletproof conditions are out there.


Can you all suggest some protein crystallization conditions (along 
with cryo conditions) for some commercially available proteins? I'm 
looking to get 6-8 different ones (and we'll just take them and see 
how it goes).  I wouldn't mind knowing unit cell parameters as well 
(just a citation works, I can have them figure it out).  I have about 
7 weeks to get everything grown and frozen and ready to go.


Any help would be greatly appreciated.  It always amazes me how 
helpful this group is.  Thank you very much.


Dave

Re: [ccp4bb] need some suggestions for crystallization

[Harry Powell]

I don't know. Is there?

On 4 Feb 2013, at Mon4 Feb 16:15, Jayashankar wrote:


Dear Powell,

Isn't it there a way to data mine the PDB or the other repository  
source for the time/duration/days of the crystals obtained.


Dr. Jayashankar Selvadurai
Hannover
Germany



On Mon, Feb 4, 2013 at 5:10 PM, Harry Powell lmb.cam.ac.uk> wrote:

Hi David

try going back to the one that started it all,* myoglobin, a recipe  
is at


http://www.rigaku.com/products/protein/recipes

(* feel free to argue about this)

On 4 Feb 2013, at Mon4 Feb 16:03, David Roberts wrote:

So, I know I say this every time I post on this board, but here it  
goes again.


I'm at an undergrad only school, and every 2 years I teach a class  
in protein crystallography.  This year I'm being super ambitious,  
and I'm going to take a class of 16 to the synchrotron for data  
collection.  It's just an 8 hour thing, to show them the entire  
process.  I'm hoping that we can collect 5-6 good data sets while  
there.


I would like them to grow their own crystals, and go collect data.  
Then we'd come back and actually do a molecular replacement  
(pretty easy/standard really).  Just to get a feel for how it works.


The protein I do research on is not one that I would push on this,  
as the crystals are hard to grow, they are very soft, and the data  
just isn't the best (resolution issues).  I do have a few that  
will work on my proteins, but I was thinking of having others in  
the class grow up classic proteins for data collection.  Obviously  
lysozyme is one, but I was wondering what other standard  
bulletproof conditions are out there.


Can you all suggest some protein crystallization conditions (along  
with cryo conditions) for some commercially available proteins?   
I'm looking to get 6-8 different ones (and we'll just take them  
and see how it goes).  I wouldn't mind knowing unit cell  
parameters as well (just a citation works, I can have them figure  
it out).  I have about 7 weeks to get everything grown and frozen  
and ready to go.


Any help would be greatly appreciated.  It always amazes me how  
helpful this group is.  Thank you very much.


Dave


Harry
--
Dr Harry Powell, MRC Laboratory of Molecular Biology, MRC Centre,  
Hills Road, Cambridge, CB2 0QH
Chairman of European Crystallographic Association SIG9  
(Crystallographic Computing)








Harry
--
Dr Harry Powell, MRC Laboratory of Molecular Biology, MRC Centre,  
Hills Road, Cambridge, CB2 0QH
Chairman of European Crystallographic Association SIG9  
(Crystallographic Computing)

Re: [ccp4bb] need some suggestions for crystallization

[Jayashankar]

Dear Powell,

Isn't it there a way to data mine the PDB or the other repository source
for the time/duration/days of the crystals obtained.

Dr. Jayashankar Selvadurai
Hannover
Germany



On Mon, Feb 4, 2013 at 5:10 PM, Harry Powell wrote:

> Hi David
>
> try going back to the one that started it all,* myoglobin, a recipe is at
>
> http://www.rigaku.com/products/protein/recipes
>
> (* feel free to argue about this)
>
> On 4 Feb 2013, at Mon4 Feb 16:03, David Roberts wrote:
>
> So, I know I say this every time I post on this board, but here it goes
> again.
>
> I'm at an undergrad only school, and every 2 years I teach a class in
> protein crystallography.  This year I'm being super ambitious, and I'm
> going to take a class of 16 to the synchrotron for data collection.  It's
> just an 8 hour thing, to show them the entire process.  I'm hoping that
> we can collect 5-6 good data sets while there.
>
> I would like them to grow their own crystals, and go collect data. Then
> we'd come back and actually do a molecular replacement (pretty
> easy/standard really).  Just to get a feel for how it works.
>
> The protein I do research on is not one that I would push on this, as the
> crystals are hard to grow, they are very soft, and the data just isn't the
> best (resolution issues).  I do have a few that will work on my proteins,
> but I was thinking of having others in the class grow up classic proteins
> for data collection.  Obviously lysozyme is one, but I was wondering what
> other standard bulletproof conditions are out there.
>
> Can you all suggest some protein crystallization conditions (along with
> cryo conditions) for some commercially available proteins?  I'm looking
> to get 6-8 different ones (and we'll just take them and see how it goes).
> I wouldn't mind knowing unit cell parameters as well (just a citation
> works, I can have them figure it out).  I have about 7 weeks to get
> everything grown and frozen and ready to go.
>
> Any help would be greatly appreciated.  It always amazes me how helpful
> this group is.  Thank you very much.
>
> Dave
>
>
> Harry
> --
> Dr Harry Powell, MRC Laboratory of Molecular Biology, MRC Centre, Hills
> Road, Cambridge, CB2 0QH
> Chairman of European Crystallographic Association SIG9 (Crystallographic
> Computing)
>
>
>
>
>

Re: [ccp4bb] need some suggestions for crystallization

[Harry Powell]

Hi David

try going back to the one that started it all,* myoglobin, a recipe  
is at


http://www.rigaku.com/products/protein/recipes

(* feel free to argue about this)

On 4 Feb 2013, at Mon4 Feb 16:03, David Roberts wrote:

So, I know I say this every time I post on this board, but here it  
goes again.


I'm at an undergrad only school, and every 2 years I teach a class  
in protein crystallography.  This year I'm being super ambitious,  
and I'm going to take a class of 16 to the synchrotron for data  
collection.  It's just an 8 hour thing, to show them the entire  
process.  I'm hoping that we can collect 5-6 good data sets while  
there.


I would like them to grow their own crystals, and go collect data.  
Then we'd come back and actually do a molecular replacement (pretty  
easy/standard really).  Just to get a feel for how it works.


The protein I do research on is not one that I would push on this,  
as the crystals are hard to grow, they are very soft, and the data  
just isn't the best (resolution issues).  I do have a few that will  
work on my proteins, but I was thinking of having others in the  
class grow up classic proteins for data collection.  Obviously  
lysozyme is one, but I was wondering what other standard  
bulletproof conditions are out there.


Can you all suggest some protein crystallization conditions (along  
with cryo conditions) for some commercially available proteins?   
I'm looking to get 6-8 different ones (and we'll just take them and  
see how it goes).  I wouldn't mind knowing unit cell parameters as  
well (just a citation works, I can have them figure it out).  I  
have about 7 weeks to get everything grown and frozen and ready to go.


Any help would be greatly appreciated.  It always amazes me how  
helpful this group is.  Thank you very much.


Dave


Harry
--
Dr Harry Powell, MRC Laboratory of Molecular Biology, MRC Centre,  
Hills Road, Cambridge, CB2 0QH
Chairman of European Crystallographic Association SIG9  
(Crystallographic Computing)

[ccp4bb] need some suggestions for crystallization

[David Roberts]

So, I know I say this every time I post on this board, but here it goes 
again.


I'm at an undergrad only school, and every 2 years I teach a class in 
protein crystallography.  This year I'm being super ambitious, and I'm 
going to take a class of 16 to the synchrotron for data collection.  
It's just an 8 hour thing, to show them the entire process.  I'm hoping 
that we can collect 5-6 good data sets while there.


I would like them to grow their own crystals, and go collect data. Then 
we'd come back and actually do a molecular replacement (pretty 
easy/standard really).  Just to get a feel for how it works.


The protein I do research on is not one that I would push on this, as 
the crystals are hard to grow, they are very soft, and the data just 
isn't the best (resolution issues).  I do have a few that will work on 
my proteins, but I was thinking of having others in the class grow up 
classic proteins for data collection.  Obviously lysozyme is one, but I 
was wondering what other standard bulletproof conditions are out there.


Can you all suggest some protein crystallization conditions (along with 
cryo conditions) for some commercially available proteins?  I'm looking 
to get 6-8 different ones (and we'll just take them and see how it 
goes).  I wouldn't mind knowing unit cell parameters as well (just a 
citation works, I can have them figure it out).  I have about 7 weeks to 
get everything grown and frozen and ready to go.


Any help would be greatly appreciated.  It always amazes me how helpful 
this group is.  Thank you very much.


Dave