Hi Kevin, Could you tell me more detail about beta-mercaptoethanol problem?
Min-Kyu | -----Original Message----- | From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of | Kevin Jin | Sent: Monday, March 12, 2012 3:32 PM | To: CCP4BB@JISCMAIL.AC.UK | Subject: Re: [ccp4bb] My protein precipitates at r.t and dissolves at 4 | oC | | I remember I saw the similar problem caused by beta-mercaptoethanol. | | | Kevin | | | On Mon, Mar 12, 2012 at 1:29 PM, Artem Evdokimov | <artem.evdoki...@gmail.com> wrote: | > Could be one of those weird behaviors displayed by detergents where | > cloud point anomalously changes with temperature... | > | > Artem | > | > On Mar 12, 2012 1:11 PM, "Min-Kyu Cho" <min-kyu....@live.com> wrote: | >> | >> I am using KPi buffer at pH 5.5, 100mM KCl, 2mM beta-mercaptoethanol, | >> 0.02% NaN3. | >> | >> Yes, I agree I should check CD melting curve to see temperature | >> preference of my protein. | >> | >> Min-Kyu | >> | >> | -----Original Message----- | >> | From: Kevin Jin [mailto:kevin...@gmail.com] | >> | Sent: Monday, March 12, 2012 11:16 AM | >> | To: Min-Kyu Cho | >> | Cc: CCP4BB@jiscmail.ac.uk | >> | Subject: Re: [ccp4bb] My protein precipitates at r.t and dissolves | >> at 4 | >> | oC | >> | | >> | Which kind of buffer you use? If it is Tris, then temperature | >> change will | >> | cause pH change. | >> | | >> | Actually, this is a good way for crystallization. | >> | | >> | Kevin | >> | | >> | On Mon, Mar 12, 2012 at 9:02 AM, Min-Kyu Cho | >> <min-kyu....@live.com> | >> wrote: | >> | > Hi all, | >> | > | >> | > I have a homotetrameric coiled-coil domain sample with 45aa per | each. | >> | > While I store this sample at 4oC, the sample looks clear w/o any | >> | > particles. But when I took out the sample to my bench at r.t, I | >> can | >> | > see there are precipitates (as stack of needle like particles) | >> at the | >> | > bottom of the tube after several hours. Interestingly, when I | >> put it | >> | > back into 4oC fridge, the precipitates disappeared and the | >> solution | >> | turned into clear again. | >> | > | >> | > Does anyone have knowledge of such behavior of any protein? I | >> | > appreciate any information related. | >> | > | >> | > Min-Kyu
