Re: [ccp4bb] Secrets of ZYMIT

Could you take a solution of it, buffer exchange/size exclusion and then perform MS fingerprinting to identify the protease if you don't have any luck with the companies? Best Dan Get Outlook for Android From: CCP4 bulletin board on beh

[ccp4bb] Secrets of ZYMIT

Hi everyone, this is Joerg Stetefeld. I would like to approach the community in a peculiar case of a specific proteolytic cleavage during crystallization attempts. Performing several crystallization screens we figured out that ZYMIT (https://www.ipcol.com/cleaners/zymit-low-foam) is the reas

Re: [ccp4bb] Optimization from needle shaped crystals

Dear prem, I am sharing my experience with needle crystal that worked for me. 1.First check that salt or a protein crystal by SDS PAGE or mass spectroscopy. 2. For improvement of morphology and size, you could try microseeding of your crushed crystal, generally needle crystal appear quickly, fur

Re: [ccp4bb] One protein, two data sets

Dear Prerana, the structures of the protein belonging to these two crystal forms are, strictly speaking, different. Exactly _how_ different they are can only be stated after solving them. You will then be able to compare _three_ structures of the protein - two in crystal form 1., one in crysta

[ccp4bb] General interest

Dear Colleagues, The recent IUCr Committee on Data Satellite Workshop at ECM32 Vienna on “Data science skills in publishing” speaker presentations, overall report and photos from the day are now available via the IUCr CommDat Public Forum weblink:- https://forums.iucr.org/viewtopic.php?f=39&t=4

Re: [ccp4bb] One protein, two data sets

I take it that the datasets are from different crystals. Are the crystallisation conditions the same? I can't spot any signs of a difference in indexing (but I may be wrong) so they look like different crystal forms. Are the processing statistics good for both? Is this a case for molecular repl

Re: [ccp4bb] Optimization from needle shaped crystals

4. Matrix microseeding. Make a seed stock from these crystals and then re-run your primary screens. https://www.ncbi.nlm.nih.gov/m/pubmed/25195878/ -- Dr David C. Briggs Senior Laboratory Research Scientist Signalling and Structural Biology Lab The Francis Crick Institute London, UK == about.me/

Re: [ccp4bb] Optimization from needle shaped crystals

I can share what has worked for my crystals : 1. You can put a grid across your condition with same or altered drop ratios. 2. You can try micro seeding. (This has given me the best results so far). 3. You can try Hampton's additive screen. On Sun, Sep 8, 2019 at 10:09 AM Prem Prakash wrote:

[ccp4bb] One protein, two data sets

Dear all, We have two data sets of a protein with the following parameters: 1. Space group P212121 a=61.0, b=100.34, c=133.23 No. of molecules in ASU - 2 2. Space group P212121 a=58.33, b=62.86, c=109.45 No. of molecules in ASU - 1 Can we use them as two different structures? Regards, Prerana #

Re: [ccp4bb] Optimization from needle shaped crystals

Dear Prem, before moving on with any optimization rounds it would be good to first confirm that these crystals are actually protein crystals, and in particular the protein of interest. For example, a Silver-stained SDS-PAGE gel can be very informative. You can run a couple of your crystalline c