Around here, I would guesstimate the success rate to be in the
5-10% range. Still, I always try it as it's so easy. I don't think
longer soak times will usually do much to increase occupancy. Instead,
try higher halide concentrations.
I also like to try monovalent cations (Rb, Cs) but h
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> -Original Message-
> From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.u
nnini
Sent: Tuesday, March 31, 2009 9:11 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Halide soaking
Hi,
it worked very nice for me in 1 out of 1 case where I tried it :-).
Very well diffracting crystals (1.8 Ang), rather small protein 20
kDa, 50 %solvent content, 1 mol/ASU. 20-30 s soak in
Somewhat to our surprise, we have found that, even when the halide
signal is too weak to solve the substructure from scratch, when you
find the sites (in our case, by using the protein model as a
"substructure" in Phaser), they can still add significant phase
information. So you can get mo
Hi,
it worked very nice for me in 1 out of 1 case where I tried it :-).
Very well diffracting crystals (1.8 Ang), rather small protein 20
kDa, 50 %solvent content, 1 mol/ASU. 20-30 s soak in 0.5 M NaBr
resulted in 6 nice ordered sites.
It was crucial for us to collect a 3 wavelength MAD da
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email: j-kell...@northwestern.edu
***
- Original Message -
From:
To:
Sent: Tuesday, March 31, 2009 1:07 PM
Subject: Re: [ccp4bb] Halide soaking
I'd like to remind folks here that if one's crystals are sturdy
I'd like to remind folks here that if one's crystals are sturdy enough to
survive halide soaking, they're *probably* also sturdy enough to live
through covalent iodination. Iodination is easy to set up and if it does
work out - one gets awesome quality derivatives with multiple sites (as
long as th
You always get the entry of Bromide into crystal by quick soaking,
because it does not require the incorporation of Bromide into the
protein. But whether the signal is good enough for phasing is another
story. You have to collect the full data set to know the answer.
Nian
2009/3/31 tat cheung che
We were on a Br-soak flurry for awhile, and when I also talked to others,
it would work about 20-30% of the time. A few years I compared a structure
with Se-Met and native with Br-soak. There were four Br's at about 30%
occupancy, and it phased fine. It wasn't as good as the Se-Met phasing,
but we
On Tuesday 31 March 2009 09:57:13 Jose Antonio Cuesta-Seijo wrote:
> Hi!
>
> Normally the cell parameters, etc change very very little. You'll
> only know if the bromides got in at the synchrotron by looking at the
> fluorescence spectrum
That won't help, normally. It only tells you that th
Hi!
Normally the cell parameters, etc change very very little. You'll
only know if the bromides got in at the synchrotron by looking at the
fluorescence spectrum and at the anomalous signal. Normally some will
make it in and some will be in ordered sites, then it becomes mostly
a question
Hi all
I am now trying to do bromide soaking, but i am not really sure does the
bromide atom enter my crystal. So is there any signs that indicate the entry of
bromide atom? e.g. does the space group, cell dimension change? or just nothing
change, and the bromide atom just get in?
Thanks very
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