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<https://aka.ms/ghei36> ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Jorg Stetefeld <jorg.stetef...@umanitoba.ca> Sent: Sunday, September 8, 2019 6:01:49 PM To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> Subject: [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 reason for a highly specific, but undesireable cleavage of protein components in our setups. Most remarkably, the cleavage site of our crystallisation target is not described in any protease database, and is highly inaccessible. According to a publication by Naschberger et al, 2014 (doi:10.1107/S2053230X14026053) the authors very nicely describe the phenomenon. They also describe a very elegant protease removal protocol, which works in our hands very well. At this point, however, I would like to know what is “behind” ZYMIT? Naschberger et al say “Zymit contains an unspecified‘protease enzyme’ as a trade secret (http://www.ipcol.com/pdfs/Zymit_msds.pdf)”. Does anyone has more insight into the secrets of ZYMIT? Our attempts to contact several vendors here in Canada/ abroad failed and a further characterisation of its nature would help us to understand this particular phenomenon of proteolytic cleavage. Thank you very much in advance- Your advise is appreciated. js Jörg Stetefeld <https://stetefeldlab.ca/> https://stetefeldlab.ca/<https://stetefeldlab.ca/> [X] ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
