hey, my information is that detecting phosphorylation via ms is non-trivial because the P groups are lost during fragmentation and prior to detection. cheers jon
-----Ursprüngliche Nachricht----- Von: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> Im Auftrag von Wim Burmeister Gesendet: Mittwoch, 15. September 2021 14:47 An: CCP4BB@JISCMAIL.AC.UK Betreff: Re: [ccp4bb] Phosphatase enzymatic assay Hello, try mass spectrometry, either after tryptic digestion into peptides or electrospray MS on the entire protein if it is not too big. Best Wim ----- Mail original ----- De: "Andrea Moretti" <andrea.more...@unige.ch> À: "CCP4BB" <CCP4BB@JISCMAIL.AC.UK> Envoyé: Mercredi 15 Septembre 2021 14:28:45 Objet: [ccp4bb] Phosphatase enzymatic assay Dear CCP4 community, sorry for the off topic question but I am struggling with enzymatic assays and I would need your help. I am trying to measure dephosphorylation of a phosphorylated kinase by a ser/thr phosphatase. So far I tried to measure it by malachite green assay, anti pSer/pThr antibody and PNPase coupled assay but nothing really worked for me. NMR is unfortunately not an option since I can't make enough of the proteins. So any hints on phosphatase assays would be helpful. Thanks! Best wishes, Andrea -- Andrea Moretti Structural Plant Biology Laboratory Department of Botany and Plant Biology Sciences III University of Geneva 30 Quai E. Ansermet 1211 Geneva Switzerland Email: andrea.more...@unige.ch Phone: +41 22 379 3029 (office) web:http://structplantbio.org ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/ -- Wim Burmeister Professor Institut de Biologie Structurale (IBS) CIBB 71 avenue des Martyrs / CS 20192 38044 Grenoble Cedex 9, FRANCE E-mail: [ mailto:wim.burmeis...@ibs.fr | wim.burmeis...@ibs.fr ] Mobile: +33 (0) 7 50 49 19 91 [ http://www.ibs.fr/research/research-groups/viral-replication-machines-group-m-jamin/team-03/article/poxvirus-replication-machinery-presentation/ | website ] Changement climatique : «Les autres combats n’ont aucun sens si celui-là est perdu» ( Aurélien Barrau ) ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/ ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
