Hi,
In addition to all that's been suggested so far: when I was facing such
problems I always tried the (commercially available) additive screens
(additive kits, whatever they are called nowadays). This means setting
up quite a few crystallisation experiments (you already have conditions
in w
Hi Saurabh,
I have also encountered a similar problem. Sometimes, though your protein
looks pure in size exclusion chromatography, but actually is not very pure
and may contain traces of partially unfolded form or some transient
oligomeric forms. In such a case, your size exclusion profile would b
Dear Saurabh
I would like to share something that worked for me. I was in a similar
situation, though my crystals weren't as beautiful as yours are. I followed
Alexander McPherson's Crystallization of Biological Macromolecules and what
ultimately worked is as follows:
I did something called iterat
Hi Saurabh
Of course you should use "rMMS" microseeding. See below
I hope it works !
Best wishes, Patrick
-- Forwarded message -
From: Patrick Shaw Stewart
Date: Mon, Sep 9, 2019 at 10:12 AM
Subject: Re: [ccp4bb] Optimization from needle shaped crystals
To: David Briggs
Cc:
Dear Saurabh,
It's well known that good-looking crystals can have poor internal order,
and therefore yield poor diffraction, and vice versa. As others have
pointed out, it's crucial to check the quality of diffraction at room
temperature (RT).
Just to add a bit to that:
There's a nice book that
s/mcl
From: CCP4 bulletin board on behalf of Saurabh Upadhyay
Sent: Sunday, March 21, 2021 1:19 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Suggestions to improve resolution of protein crystals
Dear Everton and Nukri,
Thank you for your kind reply, we
Dear Everton and Nukri,
Thank you for your kind reply, we were also speculating that there could be
some issues with cryoprotection.
We have used two cryoprotectants naming; Parabar (10312) and Paraffin Oil
from Hampton Research company. We have also used glycerol and respective
mother liquors as
Dear Saurab,
It's a shame these crystals do not diffract well - they look so good!
I assume the diffraction you saw was recorded at cryo temperatures? If so,
it is possible that you may be damaging the crystals during cryo-pretection
and cryo-cooling. Before you go too deep into changing the crysta
Dear Saurabh,
Your crystals are gorgeous!
Which cryoprotectant did you use?
Best,
Everton.
Sent from my iPhone
> On Mar 21, 2021, at 11:04 AM, Saurabh Upadhyay
> wrote:
>
>
> Dear All,
>
> I am trying to crystalize a protein, which occurs in monomeric (60kDa) and
> te
