Re: [ccp4bb] Crystal optimization

Hi 白雪慧 Initially I would try to diffract these crystals befofe any optimisation. A microfocus beam should give you some clues. You might find out that you have a super ordered crystal at 1.2 A resolution already, or you crystallized salt or a contaminant. Further, have you tried seeding? These

Re: [ccp4bb] Crystal optimization

Hi, these crystals look large enough for VMXm at Diamond Light Source. We have collected excellent data out of smaller crystals. Get in touch with me (or any other members of the team) and we can discuss a possible visit. Kind regards, Jose

Re: [ccp4bb] Crystal optimization

Hi, In addition, you might want to test additives. I would suggest the additive screen from Hampton Research. We made best experiences with neutral solvents like: ethylacetate (doi.org/10.1021/ic010790t and doi.org/10.1021/acs.inorgchem.6b00672), dioxane (doi.org/10.1021/acs.inorgchem.6b00672)

Re: [ccp4bb] Crystal optimization

Usually, you should try to push up the protein concentration, often quite a lot (30, 50, even 100 mg/ml), and decrease precipitant (might have to be really low, eg <3% PEG is not unthinkable). To get the protein up, you may need to find a new buffer solution - there are screens for this, and DS

Re: [ccp4bb] Crystal optimization

I believe someone has already mentioned matrix seeding- taking the crystals you currently have, making microseeds and then using this in a new screen (whatever your favourite initial screens may be). This can give you different conditions that will give you better crystals as you already have se

Re: [ccp4bb] Crystal Optimization

ir.com >> >> b...@ruppweb.org >> >> http://www.vienna-air.com/ >> >> - >> >> It is not your aptitude but your attitude >> >> that determines your altitude. (or your crystals) >> >> -

Re: [ccp4bb] Crystal Optimization

* > > It is not your aptitude but your attitude > > that determines your altitude. (or your crystals) > > - > > ** ** > > ** ** > > ** ** > > From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of > yybbll > > Sent: Tuesday, July 10, 2012 11:58 AM >

Re: [ccp4bb] Crystal Optimization

als) - From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of yybbll Sent: Tuesday, July 10, 2012 11:58 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Crystal Optimization Hi, In my experience, it is very very very difficult to optim

Re: [ccp4bb] Crystal Optimization

Hi, In my experience, it is very very very difficult to optimize this needle like crystal. Always give up. Good luck! Dear All; Could somebody give a nice suggestion how the following type crystal could be optimized, I almost tried everything. Crystal Image is attached Crystal condition: 20%

Re: [ccp4bb] Crystal Optimization

bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Greg Costakes Sent: Tuesday, July 10, 2012 2:18 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Crystal Optimization Have you tried multiple rounds of micro-seeding

Re: [ccp4bb] Crystal Optimization

Have you tried multiple rounds of micro-seeding? --- Greg Costakes PhD Candidate Department of Structural Biology Purdue University Hockmeyer Hall, Room 320 240 S. Martin Jischke Drive, West Lafayette, IN 47907 --

Re: [ccp4bb] Crystal Optimization

Give a list of all you have tried? JPK On Tue, Jul 10, 2012 at 12:22 PM, Muhammed bashir Khan < muhammad.bashir.k...@univie.ac.at> wrote: > Dear All; > > Could somebody give a nice suggestion how the following type crystal could > be optimized, I almost tried everything. > > Crystal Image is att

Re: [ccp4bb] Crystal Optimization: Summary

Dear All, Thank you for the replies to my post. I received more than 14 response. The summary is given below. The suggestions were - add another purification step. * Ni-column * remove His-tag * Ni-column * gel filtration if you have done that: add another step to test for better crystals

Re: [ccp4bb] Crystal Optimization

Dear Jobi, See if you can slowly increase the PEG concentration to 28 - 30% and that will be a good cryo. Since BIS Tris Propane is the buffer, I think, 28% 3350 should work fine. If the crystals crack by going straight to 28% PEG 3350, 0.1M Bis Tris Propane pH:6.5, 0.2M Potassium thiocyanate

Re: [ccp4bb] Crystal Optimization

Hi Jobi- You might want to try using drop ratios ---we have had great success with this many times. My favorite "additive screen" is using the Hampton Research Crystal Screen HT as an additive screen. I usually start by adding 5% to the well---this has often yielded good crystals where the tr

Re: [ccp4bb] Crystal Optimization

Hi Jobi, I also had some crystals that were highly sensitive to glycerol. In one case, I found that DMSO at 10-15% can cryo protect it, also the crystal could grow in the presence of 10% DMSO which essentially eliminated a soaking step. In another case, I grew the crystals in the presence of 5%

Re: [ccp4bb] Crystal Optimization

Dear Jobi, the paper of Heras and Martin 'Post-crystallization treatments for improving diffraction quality of protein crystals' is really helpful. Additionally, if you have lysines have you tried reductive methylation? Good luck, e On Wed, April 13, 2011 12:34, Bingfa Sun wrote: > Dear Jobi, >

Re: [ccp4bb] Crystal Optimization

Dear Jobi, For a crystal which is big in size(0.2-0.3mm is pretty big) while diffract poorly, dehydration (increase concentration of the precipitant slowly) is a good choice to improve diffraction, especially for those tends to crack during cryo. Also those regular optimization approaches: Add

Re: [ccp4bb] Crystal Optimization

Dear Jobi, The usual suspects are: - add another purification step. In fact I'd do that first: * Ni-column * remove His-tag * Ni-column * gel filtration if you have done that: add another step to test for better crystals - additive screens - room temperature data set to check if freezin