Hi everyone,
thx for the overwhelming response, it looks like some of us experienced the very same problem, in a nutshell, dependent on vendor/product line there are three major incridients, in common to all products is the presence of three protease sub-classes, we will perform some test runs with a couple of model targets and compile/publish a list thx again- it helped us a lot js Jörg Stetefeld <https://stetefeldlab.ca/> https://stetefeldlab.ca/<https://stetefeldlab.ca/> [X] ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Bernhard Rupp <hofkristall...@gmail.com> Sent: Monday, September 9, 2019 5:00 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Secrets of ZYMIT Well, trade secret means exactly what it says. We were equally unable to extort it from the suppliers. MS fragment search as suggested might work. Best, BR From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> On Behalf Of Jorg Stetefeld Sent: Monday, September 9, 2019 00:02 To: 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/ ________________________________ 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 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
