Re: [ccp4bb] Crystallisation textbook

I second the Acta F special issue (as an unbiased reviewer): https://journals.iucr.org/special_issues/2016/crystallization/ Best, Aaron -- Aaron D. Finke, PhD Staff Scientist, MAX IV Laboratory e-mail: aaron.fi...@maxiv.lu.se On Jul 14, 2023, at 17:19, Ho

Re: [ccp4bb] Crystallisation textbook

Dear Mike,And a review of our book in Chemistry World:-Crystal clearchemistryworld.comGreetings,John Emeritus Professor John R Helliwell DScOn 14 Jul 2023, at 16:20, John R Helliwell wrote:Dear Mike,I offer you this suggestion :-global.oup.comwhich is in the OUP and IUCr Book Series. Greetings,Jo

Re: [ccp4bb] Crystallisation textbook

Dear All, The Acta Cryst F special issue is also a very good resource (although I admit that I am biased). Can be read online here: https://journals.iucr.org/special_issues/2016/crystallization/ or ordered as a book from the IUCr h

Re: [ccp4bb] Crystallisation textbook

Macromolecular Crystallization and Crystal Perfection that John has already recommended. Briony From: CCP4 bulletin board on behalf of Hough, Michael (DLSLtd,RAL,LSCI) <69715b1ac6c0-dmarc-requ...@jiscmail.ac.uk> Date: Friday, 14 July 2023 at 15:19 To: CCP4BB@JISCMAIL.AC.UK Subject: [

Re: [ccp4bb] Crystallisation textbook

Dear Mike,I offer you this suggestion :-global.oup.comwhich is in the OUP and IUCr Book Series. Greetings,John Emeritus Professor John R Helliwell DScOn 14 Jul 2023, at 15:19, Hough, Michael (DLSLtd,RAL,LSCI) <69715b1ac6c0-dmarc-requ...@jiscmail.ac.uk> wrote: Hi all,   I'd be grateful f

Re: [ccp4bb] Crystallisation textbook

When I first started crystallization made crystal clear (by Gale Rhodes) and biomolecular crystallography (by Bernhard Rupp) were my favourites. I think the former may have a new version coming out soon. Best, Omid On Jul 14, 2023, at 08:20, Hough, Michael (DLSLtd,RAL,LSCI) <69715b1ac6c0-d

Re: [ccp4bb] Crystallisation textbook

: [ccp4bb] Crystallisation textbook Hi all, I'd be grateful for any recommendations of a good up-to-date textbook covering macromolecular crystallisation. Currently we are using the excellent book by Terese Bergfors (2009) but would be interested if there is anything more recent with discu

[ccp4bb] Crystallisation textbook

Hi all, I'd be grateful for any recommendations of a good up-to-date textbook covering macromolecular crystallisation. Currently we are using the excellent book by Terese Bergfors (2009) but would be interested if there is anything more recent with discussion around automation, micro-crystallis

[ccp4bb] Crystallisation Laboratory Manager (Senior Support Scientist) - VMXi beamline

Dear All A position to join the VMXi team at Diamond Light Source as Crystallisation Laboratory Manager is available (Closing date is March 5th) The VMXi team is looking to recruit a crystallisation laboratory manager who will be part of the beamline team and be responsible for managing crysta

Re: [ccp4bb] Crystallisation of a minority fraction monomers

Dear Sebastiaan, we indeed found something very similar with selecase (see http://www.ncbi.nlm.nih.gov/pubmed/25159620). This is a highly specific metallopeptidase, which is also a metamorphic protein that transits between several stable states, among which only the monomer is the active species. W

Re: [ccp4bb] Crystallisation of a minority fraction monomers

I just wanted to disagree with Roger's word choice, but not his argument (this is a "flame"-free response). Forget about "packing" and "packable" as there is no outside force doing the work. The molecules are just falling into a local energy minimum where favorable intra- and intermolecular in

Re: [ccp4bb] Crystallisation of a minority fraction monomers

Just for clarity: symmetry mates have been taken into account, the dimer really doesn't show up in those crystals. Seb. At Wednesday, 08-04-2015 on 15:16 Marjolein Thunnissen wrote: Hi I guess you mean that the protein is a monomer in the asymmetric unit. It is quite common for multimer

Re: [ccp4bb] Crystallisation of a minority fraction monomers

The problem with crystallization is that is selects for the least soluble, most packable species. Sometimes that works against what you would like to know. That could include oligomerization state as well as conformational state. For example, some of the allosteric carbonic anhydrases stubbornl

Re: [ccp4bb] Crystallisation of a minority fraction monomers

Sure - crystallization selects what packs, which may constitute a very minor fraction of what you set up drops with. 1DUH is an example... -Luca > On Apr 8, 2015, at 15:08, Sebastiaan Werten > wrote: > > Dear all, > > we are currently working on a protein that is known to exist in a > monome

[ccp4bb] Crystallisation of a minority fraction monomers

Dear all, we are currently working on a protein that is known to exist in a monomer-dimer equilibrium. At the high concentrations used for crystallisation assays, the dimer is predominant and the monomer practically undetectable. Nevertheless, one of the crystal forms that we have obtained co

[ccp4bb] crystallisation: sorry, sort of off topic

Dear Dean, The bigger conferences are your best bet (IUCR, ACA etc) and also all the synchrotron user meetings, which usually have a good exhibitor attendance. Cheers, Jose' Beamline I04-1 Diamond Light Source

Re: [ccp4bb] Crystallisation optimisation query

Hi everyone, Thank you for your responses I will keep on with optimisation! Cameron -Original Message- From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of CAMERON FYFE Sent: 11 December 2013 14:57 To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Crystallisation

Re: [ccp4bb] Crystallisation optimisation query

Dear Cameron, Sokalan CP7 is a maleic acid/acrylic acid copolymer manufactured by BASF. You can find some (limited) information about the chemistry of some Sokalans within the BASF web resources. We describe our tests of that polymer as a precipitant in the following paper: Grimm C et al.

[ccp4bb] Crystallisation optimisation query

Hi everyone, I have been working with a protein 182 kDa in size and the protein has been purified in 200 mM NaCl, 50 mM Tris, and pH7.5. After incubating with protease and purifying by size exclusion I have taken one of the main peaks and managed to get good looking crystals with the largest b

Re: [ccp4bb] Crystallisation below 0°C

If your precipitant is high concentration salt, there will be a significant freezing point depression (see e.g. wikipedia entry http://en.wikipedia.org/wiki/Freezing-point_depression ). I seem to remember collecting data from crystals we cooled to -15 C in the early '80s. The crystals were grown i

Re: [ccp4bb] Crystallisation below 0°C

Hi Glenn - I have nothing systematic, but I remember that transducin-alpha (1TND) was crystallized at -12 C, with 20% glycerol in the mother liquor. (The crystals grew at higher temperatures, but weren't as good.) I worked on this project briefly in grad school, and I remember that looking a

Re: [ccp4bb] Crystallisation below 0°C

: Thursday, May 30, 2013 6:26 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Crystallisation below 0°C Hello CCP4BBers, I am currently playing with some crystals that seem to enjoy lower temperatures, and I was thinking of breaking the 0°C threshold. ...

Re: [ccp4bb] Crystallisation below 0°C

Glenn Masson, We had crystals that appeared by chance at "underfined" low temparature: Maïga, A., Vera, L., Marchetti, C., Lorphelin, A., Bellanger, L., Mourier, G., Servent, D., Gilles, N. & Stura, E. A. (2013) Crystallization of recombinant Green mamba ρ-Da1a toxin during lyophilization pro

[ccp4bb] Crystallisation below 0°C

Hello CCP4BBers, I am currently playing with some crystals that seem to enjoy lower temperatures, and I was thinking of breaking the 0°C threshold. Looking for examples of this in the literature is problematic, as searching for examples in the PDB (Under the advanced search-> crystal properties->

Re: [ccp4bb] Crystallisation video

Hi, Our lab has a few videos on youtube that may be what you are looking for. http://www.youtube.com/user/JanetSmithLab See the "Setting Crystal Trays" video. -Donald On Aug 4, 2012, at 12:18 AM, Bret Church wrote: > Hi, > > Does anybody know of a video of setting up crystallisation of HEW

Re: [ccp4bb] Crystallisation video

Jim From: CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of Bret Church [bret.chu...@sydney.edu.au] Sent: Friday, August 03, 2012 11:18 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Crystallisation video Hi, Does anybody know of a video of setti

[ccp4bb] Crystallisation video

Hi, Does anybody know of a video of setting up crystallisation of HEWL or other protein straightforward to crystallise (ie setting up "manually")? Such a setup has the potential even to be popular in schools, I think. I mean I want the experiment shown, and not just a lecture on protein cryst

[ccp4bb] Crystallisation at pH 2.0

Dear all Anybody has experience in protein crystallization at pH 2.0? Conditions, buffers, temperature...? Thanks a lot Marta Dr. Marta Martínez Júlvez Departamento de Bioquímica y Biología Molecular y Celular Facultad de Ciencias Universidad de Zaragoza Pedro Cerbuna 12 50009-Zaragoza Spain

[ccp4bb] Crystallisation Construct Designer (CCD)

Dear all, After the rather unexpected positive responses on CCD over the last few days, according to our web log and the emails we got, please note that we got motivated to: 1. Establish a Help button that explains what the program is and is not for, gives some general instructions 2. E

Re: [ccp4bb] cryo-cooling, was: Re: [ccp4bb] crystallisation and mosaicity

But for some crystals flash-cooling is better at temperatures higher than in the 77 K to 100 K regime for unknown reasons. This can be one reason why flash-cooling in the gas stream occassionally is better. It is also why liquid propane worked for Raji E. and Karolin L.: they were using a

Re: [ccp4bb] cryo-cooling, was: Re: [ccp4bb] crystallisation and mosaicity

Jim, The fact that liquid propane can exist at a range of temperatures is actually a MAJOR advantage. While you must ensure that the temperature of the liquid propane is just above its own freezing point, the very high boiling point of propane ensures that there is liquid-to-solid contact wit

Re: [ccp4bb] cryo-cooling, was: Re: [ccp4bb] crystallisation and mosaicity

Jim, The fact that liquid propane can exist at a range of temperatures is actually a MAJOR advantage. While you must ensure that the temperature of the liquid propane is just above its own freezing point, the very high boiling point of propane ensures that there is liquid-to-solid contac

[ccp4bb] cryo-cooling, was: Re: [ccp4bb] crystallisation and mosaicity

I would like to point out that flash-cooling in liquid propane has the added complication that the liquid propane can have a range of temperature and still be liquid. If you use propane you may not know which temperature you are actually using. The temperature in the exposed layer of the prop

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation and mosaicity

Tommi, The question has been asked and answered not by protein crystallography, but by cyroelectron microscopy and EM freeze etch research. Even as far back as the early 1960's, people noticed that liq. N2 was really slow at cooling. Read the cyroEM work on the bacteriorhodopsin photocyc

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation and mosaicity

according to literature,see below and references http://www.px.nsls.bnl.gov/courses/papers/ZD_EG_papers.html, it is not clear that liq. propane plunged item would cool faster. (whilst i havent tested this)... Would anyone have actual experimental data with protein crystals on the hyperquenchin

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation and mosaicity

I think the important thing here is that liquid nitrogen in the lab tends to be exactly at its boiling point, since the temperature is maintained by continuously boiling off some of the N2. This means the only mechanism for heat absorption is through vaporization, depending on the latent heat of

Re: [ccp4bb] crystallisation and mosaicity

ers on the subject as well as J. Appl. Cryst. (2006). 39, 244-251; >> Methods 34 (2004) 415-423 and Acta Cryst. (2002). D58, 459-471, to name >> a few. >> Jeff >> >> -Original Message- >> From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of >

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation and mosaicity

yes you are right, but I assumed if people see a cloud of condensed fog over their LN2 bath they should remove that by a) filling up the bowl completely e.g. some LN2 drips out of the bowl b) blow the fog away before you dip I think the original poster meant the relatively low heat conduction of

[ccp4bb] Fwd: [ccp4bb] crystallisation and mosaicity

Hi Charlie, yes you are right, but I assumed if people see a cloud of condensed fog over their LN2 bath they should remove that by a) filling up the bowl completely e.g. some LN2 drips out of the bowl b) blow the fog away before you dip True; this has been demonstrated quite rigorously:

Re: [ccp4bb] crystallisation and mosaicity

[EMAIL PROTECTED] On Behalf Of Paul Paukstelis Sent: Tuesday, June 03, 2008 11:59 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation and mosaicity I have also observed this. In my cases I have also been able to get away with using less cryoprotectant when freezing in the cryostr

Re: [ccp4bb] crystallisation and mosaicity

nd Acta Cryst. (2002). D58, 459-471, to name a few. Jeff -Original Message- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Paul Paukstelis Sent: Tuesday, June 03, 2008 11:59 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation and mosaicity I have also

Re: [ccp4bb] crystallisation

Dear All Thank you for your suggestions. I got many suggestions from you. Due to my time constraint, I'm not writing summary, but some replies for your suggestions to my previous query. I have not mentioned fully what I tried to do better diffraction. I'm listing them here which were mentioned b

Re: [ccp4bb] crystallisation and mosaicity

ne 03, 2008 11:59 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation and mosaicity I have also observed this. In my cases I have also been able to get away with using less cryoprotectant when freezing in the cryostream. --paul Daniel Pomeranz Krummel wrote: > Dear Sajid, &g

Re: [ccp4bb] crystallisation and mosaicity

I have also observed this. In my cases I have also been able to get away with using less cryoprotectant when freezing in the cryostream. --paul Daniel Pomeranz Krummel wrote: Dear Sajid, I have observed a consistent reduction in mosaicity (from approx. 0.95 to 0.45) for one case by freezing t

Re: [ccp4bb] crystallisation and mosaicity

Dear Sajid, I have observed a consistent reduction in mosaicity (from approx. 0.95 to 0.45) for one case by freezing the crystals in a cryostream (N2 vapour) as opposed to submerging in liquid nitrogen. Have others observed this? Daniel > > Dear All > > My protein size is ~30kD and crystallizes

Re: [ccp4bb] crystallisation

ition of dimethyl arsenic (from the > cacodylate) to some of your cysteines? > Also, 10% glyercol sounds quite low for reproducibly good > freezing (at least in my experience). > Phoebe > > Original message > >Date: Mon, 2 Jun 2008 17:04:38 +0100 > >From: sajid akt

Re: [ccp4bb] crystallisation

? Also, 10% glyercol sounds quite low for reproducibly good freezing (at least in my experience). Phoebe Original message Date: Mon, 2 Jun 2008 17:04:38 +0100 From: sajid akthar <[EMAIL PROTECTED]> Subject: [ccp4bb] crystallisation To: CCP4BB@JISCMAIL.AC.UK Dear All My p

Re: [ccp4bb] crystallisation

Jun 2008 17:04:38 +0100 >From: sajid akthar <[EMAIL PROTECTED]> >Subject: [ccp4bb] crystallisation >To: CCP4BB@JISCMAIL.AC.UK > > >Dear All > >My protein size is ~30kD and crystallizes with >19%Peg3350, 0.2M Nacl, and 0.1M Na Cacodylate buffer. > >Please

Re: [ccp4bb] crystallisation

I summarized most of the points in the CCP4 wiki at Hope nobody minds! Could future contributions please be added there? - thanks! Kay -- Kay Diederichs http://strucbio.biologie.uni-konst

Re: [ccp4bb] crystallisation

Hi Sajid, congratulations to your nice looking crystals. I'm not sure if someone already sugested to mount smaller crystals ? You write they need 4-5 days, I assume you can already see them earlier before they reach that size. Am I roughly right in the assumption that these crystals are about

Re: [ccp4bb] crystallisation

Hello Sajid, Is you crystal this mosaic in all three directions? It could be that with the longer rods, that your mosaicity is high in one direction but not so much in the other two directions. In this case I'd guess shooting down the length of the xtal might result in increased mosaicity. Thu

Re: [ccp4bb] crystallisation

Hi Sajid, with my very (!) limited experience I can recommend trying to co-crystallise with your cryo-buffer at very low concentration. It helped me (once!) to get diffraction going from about 2.7 to 1.5 ... oh, and it got rid of some nasty twinning problem as well. I guess the thinking is that if

Re: [ccp4bb] crystallisation

Hi Sajid, Just a simple test for your problem: Incubate your crystal longer in your cryo/stabilization solution. This helps sometimes to lower mosaicity. Of course, you can also try co-crystallization with glycerol (2%, 5% or 10%). Good Luck, christian sajid akthar wrote: Dear All My protein

Re: [ccp4bb] crystallisation

sajid akthar wrote: Dear All My protein size is ~30kD and crystallizes with 19%Peg3350, 0.2M Nacl, and 0.1M Na Cacodylate buffer. Please refer the attached crystal image with this. The crystal looks like good enough for home source. These crystals appears in 4-5 days at room temp. Sometimes I'

Re: [ccp4bb] crystallisation

What is the room temperature diffraction like? If that's good, you might like to optimize cryo. If I am not mistaken, PEG 3350 has been reported to cryoprotect at 20-35% with or without 5-10% glycerol. You might also like to try including some glycerol in the mother liquor--it may better crystal to

[ccp4bb] crystallisation robot

YSTAL Sent: 14 April 2008 22:10 To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot Hi, Does anyone have information about how long it takes to set up a 96-well tray for the crystallization robots available? Besides cost per tray and maintenance cost, ano

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

ary 18, 2008 2:20 AM To: <mailto:CCP4BB@JISCMAIL.AC.UK>CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Fwd: [ccp4bb] crystallisation robot One thing that people often overlook is that quite a lot of protein can be lost by denaturation on the surface of the drop. This is more significant for smaller drops.

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

EMAIL PROTECTED] ** ** Headington, Oxford OX3 7BNhttp://www.oppf.ox.ac.uk ** Original message Date: Mon, 14 Apr 2008 17:10:26 -0400 From: JOE CRYSTAL <[EMAIL PROTECTED]> Subject: Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot To: CCP4BB@JISCMAIL.AC.UK

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

Lisa A. Nagy, Ph.D. > University of Alabama-Birmingham > [EMAIL PROTECTED] > > -Original Message- > From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On > Behalf Of > Patrick Shaw Stewart > Sent: Friday, January 18, 2008 2:20 AM > To: CCP

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

Thanks Leo for your introduction. However, the link is broken. Please look at the following page. http://pfweis.kek.jp/protein/Robot/PXS/index-e.html Best regards, Yusuke --- Yusuke Yamada, Ph.D. Photon Factory High Energy Accelerator Research Organi

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

B@JISCMAIL.AC.UK *Subject:* Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot Hi, Does anyone have information about how long it takes to set up a 96-well tray for the crystallization robots available? Besides cost per tray and maintenance cost, another important feature we consider is the time

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

Dear Joe -- Does anyone have information about how long it takes to set up a 96- well tray for the crystallization robots available? Besides cost per tray and maintenance cost, another important feature we consider is the time for setting up a 96-well tray. It is an important factor sinc

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

board [mailto:[EMAIL PROTECTED] On Behalf Of JOE CRYSTAL Sent: Monday, April 14, 2008 5:10 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot Hi, Does anyone have information about how long it takes to set up a 96-well tray for the crystallization robots

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

CP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot Hi, Does anyone have information about how long it takes to set up a 96-well tray for the crystallization robots available? Besides cost per tray and maintenance cost, another important feature we consider is the time for se

Re: [ccp4bb] crystallisation robot

P4BB@JISCMAIL.AC.UK <mailto:CCP4BB@JISCMAIL.AC.UK> Subject: [ccp4bb] Fwd: [ccp4bb] crystallisation robot One thing that people often overlook is that quite a lot of protein can be lost by denaturation on the surface of the drop. This is more significant for smaller d

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

gt; -- > > > Lisa A. Nagy, Ph.D. > > > University of Alabama-Birmingham > > > [EMAIL PROTECTED] > > > > > > -Original Message- > > > From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of > > > Patrick Shaw Stewart >

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

ilto:[EMAIL PROTECTED] On Behalf Of Patrick Shaw Stewart Sent: Friday, January 18, 2008 2:20 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Fwd: [ccp4bb] crystallisation robot One thing that people often overlook is that quite a lot of protein can be lost by denaturation on the surface of the drop. T

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

aw Stewart > Sent: Friday, January 18, 2008 2:20 AM > To: CCP4BB@JISCMAIL.AC.UK > Subject: [ccp4bb] Fwd: [ccp4bb] crystallisation robot > > One thing that people often overlook is that quite a lot of protein > can be lost by denaturation on the surface of the drop. This is more > s

Re: [ccp4bb] Fwd: [ccp4bb] crystallisation robot

ECTED] -Original Message- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Patrick Shaw Stewart Sent: Friday, January 18, 2008 2:20 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Fwd: [ccp4bb] crystallisation robot One thing that people often overlook is that quite a lot of protei

[ccp4bb] Fwd: [ccp4bb] crystallisation robot

___ > > > From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of > [EMAIL PROTECTED] > Sent: Wednesday, January 16, 2008 8:31 PM > To: CCP4BB@JISCMAIL.AC.UK > Subject: Re: [ccp4bb] crystallisation robot > > > > > > Once upon a time I worked in a

Re: [ccp4bb] crystallisation robot

ifferent, the end point is not, and this can significantly affect the outcome of the experiment. Mark -Original Message- From: Oganesyan, Vaheh <[EMAIL PROTECTED]> To: CCP4BB@JISCMAIL.AC.UK Sent: Thu, 17 Jan 2008 7:40 am Subject: Re: [ccp4bb] crystallisation robot

Re: [ccp4bb] crystallisation robot

heh Sent: Thursday, January 17, 2008 8:41 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation robot Mark, What was the state of the larger drops when tiny counterparts had crystals? My guess - they all precipitated. I'm trying to understand why some proteins or some condi

Re: [ccp4bb] crystallisation robot

January 16, 2008 8:31 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation robot Once upon a time I worked in a group that was interested in developing crystallization in microfluidics. This was before the time that Fluidigm existed and we had not heard of crystallization with

Re: [ccp4bb] crystallisation robot

s not a flame, even if heat might be a good thing on a night with > temperatures predicted far below 0F. > > ?:-) > > Mark > > > > > > > > -Original Message- > From: Anastassis Perrakis <[EMAIL PROTECTED]> > To: CCP4BB@JISCMAIL.AC.UK > Sent: W

Re: [ccp4bb] crystallisation robot

CCP4BB@JISCMAIL.AC.UK Sent: Wed, 16 Jan 2008 6:17 am Subject: Re: [ccp4bb] crystallisation robot > Oryxnano 50+50 nL? >? > Demetres? >? ? Which, indirectly, brings up an interesting (but not relevant to the Oryx) question.? ? Nucleation is a process that does have a stocha

Re: [ccp4bb] crystallisation robot

CCP4BB@JISCMAIL.AC.UK Sent: Wed, 16 Jan 2008 6:17 am Subject: Re: [ccp4bb] crystallisation robot > Oryxnano 50+50 nL? >? > Demetres? >? ? Which, indirectly, brings up an interesting (but not relevant to the Oryx) question.? ? Nucleation is a process that does have a stocha

Re: [ccp4bb] crystallisation robot

Message- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Oganesyan, Vaheh Sent: Wednesday, January 16, 2008 9:06 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation robot ... Once in a while I get large crystal that can be used for data collection, but in most of

Re: [ccp4bb] crystallisation robot

Vaheh, If I'm optimising in 24 well trays after a hit in 96 I generally use sitting drop trays (cryschem) as a first step and try different ratios of protein:mother liquor first rather than changing the protein concentration. I know that sometimes the crystals stick to the plastic but I use th

Re: [ccp4bb] crystallisation robot

Original Message- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Anastassis Perrakis Sent: Wednesday, January 16, 2008 8:23 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation robot > More recently, I've looked at all of the crystallization robot

Re: [ccp4bb] crystallisation robot - volume?

Regarding stochastic processes: to increase the chances of nucleation one would like to have protein in the right nucleation zone for a certain period of time. If the drop becomes too small, the ratio evaporation surface/drop volume changes. So, the nucleation zone might be reached within the

Re: [ccp4bb] crystallisation robot

More recently, I've looked at all of the crystallization robot vendors. For single lab users, all of the systems work well. Systems like the Hydra or Mosquito are less automatic, but provide the basic functions for crystallization trial setup. For more of a user facility with a large number of

Re: [ccp4bb] crystallisation robot

Oryxnano 50+50 nL Demetres Which, indirectly, brings up an interesting (but not relevant to the Oryx) question. Nucleation is a process that does have a stochastic aspect. Thus, one could argue that compromising to 200-300 nl might be better than either extremes of 50nl (too small volum

Re: [ccp4bb] crystallisation robot

In our original work on a prototype, we used the Cartesian technology. We were able to dispense 10nL+10nL drops with a large range of viscosities without difficulty. The main issue was to wash the tips after use to prevent clogging. That was with a system that could dispense 1nL droplets. The prese

Re: [ccp4bb] crystallisation robot

Oryxnano 50+50 nL Demetres David Briggs wrote: I'll defend the honour of the phoenix... (again) Bernhard Rupp 100+100 nl Dave Briggs (and all users at Univ of Manchester, UK) 100+100nl Others.. Only time we have ANY problems is when the nano dispensing tip gets clogged. Often a good wa

Re: [ccp4bb] crystallisation robot

I'll defend the honour of the phoenix... (again) Bernhard Rupp 100+100 nl Dave Briggs (and all users at Univ of Manchester, UK) 100+100nl Others.. Only time we have ANY problems is when the nano dispensing tip gets clogged. Often a good wash whilst still on the machine will clear the blockage. D

Re: [ccp4bb] crystallisation robot

: Wednesday, January 16, 2008 1:53 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation robot Dear Alexey, being involved in the development of the 'fixed needles + a few' robots and 96-well plates early on, I wonder about your bad experiences. You seem to say that t

Re: [ccp4bb] crystallisation robot

y, January 14, 2008 9:36 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation robot we have recently tried the new lid mentioned below. It works very well, Mosquito makes 3 hanging drops 96 well plate in 3-4 minutes! The film is very easy to handle and it is very transparent. The pri

Re: [ccp4bb] crystallisation robot

- From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Shirley Roberts Sent: Friday, January 11, 2008 7:25 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] crystallisation robot I sent this personal reply on wednesday to Madhavi Nalam's original question. In the light of r

Re: [ccp4bb] crystallisation robot

Dear all - Under the BIOXHIT home page, http://www.bioxhit.org you can navigate to "Section 1" or "HTP Crystallization", or simply follow the link below: http://icarus.embl-hamburg.de/bioxhit/bioXHITSection1.jsp It has quite a lot of data on what people use, available facilities, and testin

Re: [ccp4bb] crystallisation robot

Thanks to all who replied to my query. Madhavi

Re: [ccp4bb] crystallisation robot

I sent this personal reply on wednesday to Madhavi Nalam's original question. In the light of recent comments I'm posting it to the BB to support TTP labtech and Mosquito! Some of the points have been made by others now, excuse the repeats! We at YSBL have been very happy with our mosquito. We

Re: [ccp4bb] Crystallisation of a extremly soluble protein

Hi Sabine, I vaguely recall that an old colleague had used a mini detergent screen (as additive) to solve a problem such as yours. It may be worth setting up a plate or two to screen a few conditions before you embark into more molbio. You can use the 96 well plates with multiple wells (Corning sel

Re: [ccp4bb] Crystallisation of a extremly soluble protein

Hello Sabine, Maybe this is an option: Structure, Vol 14, 1617-1622, November 2006 Lysine Methylation as a Routine Rescue Strategy for Protein Crystallization best regards & good luck Christian Schneider Sabine wrote: > > Hi everyone, > > > > I am trying to crystallise an extremely soluble

Re: [ccp4bb] Crystallisation of a extremly soluble protein

: Schneider Sabine <[EMAIL PROTECTED]> Reply-To: Schneider Sabine <[EMAIL PROTECTED]> Date: Thu, 22 Feb 2007 14:08:44 - To: Conversation: Crystallisation of a extremly soluble protein Subject: [ccp4bb] Crystallisation of a extremly soluble protein Hi everyone, I am trying to crystallise

Re: [ccp4bb] Crystallisation of a extremly soluble protein

Hello, Sabine. I believe the technique that Kendall is referring to is Surface Entropy Reduction, which involves replacing large, highly entropic residues on the surface of the protein with lower entropy residues to reduce the entropy shield and facilitate crystallization. Traditionally, the

Re: [ccp4bb] Crystallisation of a extremly soluble protein

Sabine, There are protocols to modify surface residues that can help with crystallization, and make the protein less soluble. Unfortunately, I¹m drawing a blank on the details. I remember someone in Andrzej Joachimiak¹s group was working on this as a rescue approach for the structural genomics pip

[ccp4bb] Crystallisation of a extremly soluble protein

Hi everyone, I am trying to crystallise an extremely soluble and charged protein. It is ~30kDa and has an estimated PI of 5.2 and theoretical charge over pH range 4-10 from + 24 to -29. It is still happy at a concentration of 190mg/ml and fully reconstituted with its ligand. I have tried h

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