Hi Kevin,
Could you tell me more detail about beta-mercaptoethanol problem?
Min-Kyu
| -----Original Message-----
| From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of
| Kevin Jin
| Sent: Monday, March 12, 2012 3:32 PM
| To: [log in to unmask]
| 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
| <[log in to unmask]> 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" <[log in to unmask]> 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:[log in to unmask]]
| >> | Sent: Monday, March 12, 2012 11:16 AM
| >> | To: Min-Kyu Cho
| >> | Cc: [log in to unmask]
| >> | 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
| >> <[log in to unmask]>
| >> 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
|