ccp...@jiscmail.ac.uk] *On Behalf Of
> *Jürgen
> Bosch
> *Sent:* Tuesday, October 26, 2010 5:46 PM
>
> *To:* CCP4BB@JISCMAIL.AC.UK
> *Subject:* Re: [ccp4bb] Help with Optimizing Crystals
>
>
>
>
>
>
> Hi.
>
> Here is some additional information.
>
ss and/or purify your protein in the presence of these compounds.
Good Luck!
annie
From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Jürgen
Bosch
Sent: Tuesday, October 26, 2010 5:46 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Help with Optimizing Crystals
Hi.
Her
Janet Newman wrote a review on improving diffraction a few years back:
http://dx.doi.org/10.1107/S0907444905032130
it is open access.
Probably the most underappreciated aspect of diffraction is the purity of
the protien. This is because "impurities" like slightly misfolded versions
of your "nativ
>
> Hi.
>
> Here is some additional information.
>
> 1. The purification method that I used included Ni, tag cleavage, and SEC as
> a final step. I have tried samples from three different purification batches
> that range in purity, and even the batch with the worst purity seems to
> produ
Hi.
Here is some additional information.
1. The purification method that I used included Ni, tag cleavage, and SEC
as a final step. I have tried samples from three different purification
batches that range in purity, and even the batch with the worst purity seems
to produce crystals.
2. The pr
Seeding! Make seeds, rescreen with seeds. Look in many former ccp4bb posts for
references about this.
Jacob
- Original Message -
From: Jürgen Bosch
To: CCP4BB@JISCMAIL.AC.UK
Sent: Tuesday, October 26, 2010 3:47 PM
Subject: Re: [ccp4bb] Help with Optimizing Crystals
You
You did check on a gel that they are indeed your protein ?
If you have sufficient amounts available try digesting it with various
proteases and see if you can identify a stable fragment.
A less radical approach, which might not be accessible to you, you could screen
your protein for alternative
First piece of advice I have is to shove them in the beam and see what
happens. A few days ago we got high-resolution data from crystals that are
shaped like eggs. No edges on them whatsoever. In the past, saucer-shaped
crystals diffracted to 2A whereas their hexagonal 'perfect' cousins (grown
from
Hi Matt,
You'll probably get many different answers to a question like this, but what I
would do is go back to your protein and make different constructs; chop off
termini, surface mutations etc, maybe cleave off the tag. Of course more
screening and optimization might work, but my sense is tha
Hello.
I have obtained disk shaped crystals of a protein that I am working on. I
got hits in about 10 different conditions, with a few common precipitants
and pHs, and I have optimized two conditions so far. In the optimized
conditions, the crystals appear overnight, usually surrounded by or hid
10 matches
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