I have used a copper solution that worked well. The details can be found in this paper: Nucleic Acids Res. 2004 Sep 30;32(17):5192-7. PMID: 15459288 PMCID: PMC521666
Yingyun Liu ----- Original Message ----- From: Roger Rowlett <rrowl...@colgate.edu> Date: Thursday, February 28, 2013 9:52 am Subject: Re: [ccp4bb] disulfide engineering To: CCP4BB@JISCMAIL.AC.UK > In the literature, you can find examples of air oxidation, oxidized > glutathione (alone), mixture of reduced and oxidized glutathione, and > hydrogen peroxide. The correct concentrations have to be found > empirically. We are just now mushing through this with an engineered > disulfide variant. Air oxidation partially worked for us--one of the > engineered disulfides worked, one did not. > > _______________________________________ > Roger S. Rowlett > Gordon & Dorothy Kline Professor > Department of Chemistry > Colgate University > 13 Oak Drive > Hamilton, NY 13346 > > tel: (315)-228-7245 > ofc: (315)-228-7395 > fax: (315)-228-7935 > email: rrowl...@colgate.edu > > On 2/28/2013 7:55 AM, Clemens Grimm wrote: > >>Along these lines, what reagents do people use to promote disuflide > bonds, > >>i.e., the "anti-DTT?" > > > >Glutathione (red) + Glutathione (ox), redox potential is adjusted by > varying the ratio. > > > >Best, > >Clemens > > > > > >> > >>JPK > >> > >>On Thu, Feb 28, 2013 at 2:06 AM, David Briggs > <drdavidcbri...@gmail.com>wrote: > >> > >>>You might want to try "Disulfide by design" > >>> > >>> > >>> > >>>Cheers > >>> > >>>Dave > >>>On Feb 28, 2013 6:55 AM, "Careina Edgooms" <careinaedgo...@yahoo.com> > >>>wrote: > >>> > >>>>Dear CCP4 members > >>>> > >>>>I wish to engineer a disulfide bond at the dimer interface of a > protein I > >>>>am working with. Does anyone know of any available software to > assist with > >>>>this? > >>>> > >>>>Best > >>>>Careina > >>>> > >>> > >> > >> > >>-- > >>******************************************* > >>Jacob Pearson Keller, PhD > >>Postdoctoral Associate > >>HHMI Janelia Farms Research Campus > >>email: j-kell...@northwestern.edu > >>******************************************* > >> > > > > > > > >-------------------------------------------------- > >Dr. Clemens Grimm > >Institut für Biochemie > >Biozentrum der Universität Würzburg > >Am Hubland > >D-97074 Würzburg > >Germany > >e-mail: clemens.gr...@biozentrum.uni-wuerzburg.de > >phone : +49 0931 31 84031 > >-------------------------------------------------
