Re: [ccp4bb] 图片1

Hi All, There is a bit of information in the crystallization conditions that I would like to comment on. Crystals grow in a variety of conditions, including one that contains Na-Cacodylate buffer (#3 in the list). When this is the case, the cacodylate-containing conditions should be dropped from fu

Re: [ccp4bb] 图片1

Hi there, all good suggestions to optimize crystals, here are few other things to try: 1. Spin down your protein sample in a centrifuge prior to setting up trays to remove protein aggregates.2. Incubate trays at lower temperatures (for example if crystals grow at RT, try 15 and 4 C).3. Try stre

Re: [ccp4bb] 图片1

> > It is pretty normal (actually, pretty good) to get crystals within 3 days > under room temperature Why work with conditions that take 3 days for crystals to grow when you can, very likely, find conditions that grow in one day by using seeding ! ? (I'm simplifying slightly - they may take 3 d

Re: [ccp4bb] 图片1

First of all, you need to verify these crystals are protein. To do so, use a loop to fish some crystals and take a shot to see the reflection pattern, a lazy way! If they are protein, then start considering the optimization. No matter which kind of kits you have used, they are just matrices of dif

Re: [ccp4bb] 图片1

Hi ZB I . . . . removed the primary crystal condition I'm not sure exactly what procedure you are using. You should suspend the crushed crystals in the original crystallization condition and add a small amount (about 15% of the drop) to every drop in a random screen eg Crystal Screen, Index, JC

Re: [ccp4bb] 图片1

This could be a good case for VMXm at Diamond Light Source. https://www.diamond.ac.uk/Instruments/Mx/VMXm.html Email adam.craws...@diamond.ac.uk or anna.war...@diamond.ac.uk, I am sure they’d be happy help/advise. Chris Fr

Re: [ccp4bb] 图片1

Hi, to me it looks like 2 things are happening in the drops, to many nuclei and on these initial nuclei new crystals appear. This may indicate that your protein is not that soluble to start with and that there may be some protein aggregation going on in your protein stock solution. Diluting the

Re: [ccp4bb] 图片1

Hi, What size are these crystals? Chances are, you could take them as they are (with cryo-protection and cryo-cooling, of course) to a synchrotron beamline with a small beam. Also, what is the PI of the protein? If crystals grow at both pH 5.5 and 8.5, perhaps you could set up a grid with pH in one

Re: [ccp4bb] 图片1

Hi, Are these initial crystals or have you already attempted some optimization? >From the crystallization condition, it appears these are your first crystals. If so, you could try the following: 1) screen around this condition for each component separately or together (coarse grid and fine scree

Re: [ccp4bb] 图片1

If you haven't already, you should definitely try MMS microseeding (adding seed stock to a random screen). There’s a good chance you might be able to get new hits in different conditions. More info: https://www.douglas.co.uk/mms.htm Best regards, Stefan *Stefan Kolek* Douglas Instruments Ltd D

Re: [ccp4bb] 图片1

Hello, I agree that it looks difficult. I would try lowering the protein concentration with that condition. Seeding springs to mind. What is in the purification buffer? Best wishes, Jon Cooper. jon.b.coo...@protonmail.com Sent from Proton Mail Android Original Message On 05/1