Did anyone suggest adding any known ligand? If your protein happens to
bind some compound/peptide/DNA/RNA/whatever, including that other partner
could dramatically change it's solution properties and be an easy fix for
your handling/formulation issues.
Good luck!
On Fri, Jul 14, 2017 at 6:33 AM,
Hi Chris,
In your Ni-NTA buffer, the total [Na+] could be greater than 530mM after
titration. Likely, your protein likes higher concentration salt for surface
charge stabilization. Adding Glycerol may further modify the charge
distribution and make the protein happy in the solution. For further
ve
HEPES pH 7.5, 10% glycerol)
works. It should work well with crystallization.
Cheers,
Chun
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Chris Fage
Sent: Friday, July 14, 2017 3:33 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Protein rapidly precipitates when off ice
: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Protein rapidly precipitates when off ice
Dear All,
Thank you for the many suggestions. After sending my first message to the BB, I
tried exchanging the sample into buffer containing 5 mM EDTA and also into
buffer at pH 9.0 (using BICINE). Neither of these
Dear All,
Thank you for the many suggestions. After sending my first message to the
BB, I tried exchanging the sample into buffer containing 5 mM EDTA and also
into buffer at pH 9.0 (using BICINE). Neither of these appear to have
helped the instability--precipitation still occurred within ~1 min o
ripts.iucr.org/cgi-bin/paper?dz5020
>
> HTH,
>
> Dave
>
> --
> Dr David C Briggs
> Hohenester Lab
> Department of Life Sciences
> Imperial College London
> UK
> http://about.me/david_briggs
>
>
> From: CCP4 bulletin bo
t;> outlined...
>>
>> http://scripts.iucr.org/cgi-bin/paper?dz5020
>>
>> HTH,
>>
>> Dave
>>
>> --
>> Dr David C Briggs
>> Hohenester Lab
>> Department of Life Sciences
>> Imperial College London
>> UK
>> http://about
_
> From: CCP4 bulletin board on behalf of Chris Fage
>
> Sent: Thursday, July 13, 2017 11:40:34 PM
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: [ccp4bb] Protein rapidly precipitates when off ice
>
> Dear CCP4BB Community,
>
> This week, I purified a nicely overexp
College London
UK
http://about.me/david_briggs
From: CCP4 bulletin board on behalf of Chris Fage
Sent: Thursday, July 13, 2017 11:40:34 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Protein rapidly precipitates when off ice
Dear CCP4BB Community,
This we
Dear CCP4BB Community,
This week, I purified a nicely overexpressing protein by Ni-NTA followed by
gel filtration. In a 4 C centrifuge, I concentrated my gel filtration
fractions to ~1 mL, transferred the spin filter to ice, and then collected
2 uL for measurement on the Nanodrop. Sadly, the prote
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