[ccp4bb] AW: [ccp4bb] [ccp4bb] protein precipitation reg

products -Ursprüngliche Nachricht- Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] Im Auftrag von Mark Wilson Gesendet: Mittwoch, 29. März 2017 22:12 An: CCP4BB@JISCMAIL.AC.UK Betreff: Re: [ccp4bb] [ccp4bb] protein precipitation reg I heartily concur with Craig. Tris can

[ccp4bb] AW: [ccp4bb] [ccp4bb] protein precipitation reg

IL.AC.UK Betreff: Re: [ccp4bb] [ccp4bb] protein precipitation reg Since I’m in full-on cranky old biochemist mode now, I think that vitriol is an old name for sulfuric acid. > On Mar 29, 2017, at 8:40 PM, Keller, Jacob <kell...@janelia.hhmi.org> wrote: > >> And if we are going

Re: [ccp4bb] [ccp4bb] protein precipitation reg

Since I’m in full-on cranky old biochemist mode now, I think that vitriol is an old name for sulfuric acid. > On Mar 29, 2017, at 8:40 PM, Keller, Jacob wrote: > >> And if we are going to pour scorn and vitriol on Tris, why not mention its >> large dpKa/dT of 0.03 pH

Re: [ccp4bb] [ccp4bb] protein precipitation reg

>And if we are going to pour scorn and vitriol on Tris, why not mention its >large dpKa/dT of 0.03 pH units/deg ? Hah! That's what many people are doing when they make buffers: pouring vitriol (HCl) on TRIS! I prefer to pour concentrated HEPES, and get two buffers without adding any extra Cl-.

Re: [ccp4bb] [ccp4bb] protein precipitation reg

] On Behalf Of CRAIG A BINGMAN Sent: Thursday, 30 March 2017 8:54 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] [ccp4bb] protein precipitation reg > On Mar 29, 2017, at 4:15 PM, Chun Luo <c...@accelagen.com> wrote: > > In addition to price, the prevalence of Ni purification may be

Re: [ccp4bb] [ccp4bb] protein precipitation reg

> On Mar 29, 2017, at 4:15 PM, Chun Luo wrote: > > In addition to price, the prevalence of Ni purification may be another reason > for Tris popularity. Some His-tagged constructs don't bind to Ni well in > HEPES. I wonder if anyone has similar experience or comments. —Chun

Re: [ccp4bb] [ccp4bb] protein precipitation reg

@JISCMAIL.AC.UK] On Behalf Of Mark Wilson Sent: Wednesday, March 29, 2017 1:12 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] [ccp4bb] protein precipitation reg I heartily concur with Craig. Tris can be a dangerous buffer for many reasons, including those listed below. In addition, as a primary

Re: [ccp4bb] [ccp4bb] protein precipitation reg

I heartily concur with Craig. Tris can be a dangerous buffer for many reasons, including those listed below. In addition, as a primary amine, it can complicate work with metalloproteins and has moderate nucleophilicity. There is almost always a better buffer choice than Tris. Best regards, Mark

Re: [ccp4bb] [ccp4bb] protein precipitation reg

There are almost always better choices than Tris buffer. Mo Cleland used to call it “Trash” buffer. He is no longer with us, but today I will happily carry that flag in his honor. Tris may show up in your crystal structure, especially at carbohydrate binding sites. Tris may be a surprisingly