tion
Good luck
Yury
--
*From:* CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of anita p
[crystals...@gmail.com]
*Sent:* Friday, August 26, 2011 3:03 AM
*To:* CCP4BB@JISCMAIL.AC.UK
*Subject:* [ccp4bb] Protein aggregation and crystallization
Hi All,
I am working on a protein which ha
From:* CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of anita p
>> [crystals...@gmail.com]
>> *Sent:* Friday, August 26, 2011 3:03 AM
>> *To:* CCP4BB@JISCMAIL.AC.UK
>> *Subject:* [ccp4bb] Protein aggregation and crystallization
>>
>> Hi All,
>>
1 3:03 AM
> *To:* CCP4BB@JISCMAIL.AC.UK
> *Subject:* [ccp4bb] Protein aggregation and crystallization
>
> Hi All,
> I am working on a protein which has a membrane spanning region and as
> cytosolic domain.I have made various deletion constructs of the protein, so
> that I can have
Hi,
I do have 10% glycerol in my buffers, and still the constructs come in the
void volume.
and I have sarkosyl in the lysis buffer. but none in the elution or dialysis
buffer. So do I still need detergents please suggest.
reg.
Anita
On Sat, Aug 27, 2011 at 12:13 AM, Pius Padayatti wrote:
>
Hi All,
I am working on a protein which has a membrane spanning region and as
cytosolic domain.I have made various deletion constructs of the protein, so
that I can have a crystallizable fragment. There is no homologues mentioned
in the pdb for this protein.
All of these constructs are purified
ccept the risks in doing so.
--- On Wed, 3/23/11, Mark J van Raaij wrote:
From: Mark J van Raaij
Subject: Re: [ccp4bb] protein aggregation
To: CCP4BB@JISCMAIL.AC.UK
Date: Wednesday, March 23, 2011, 2:03 PM
- try limited proteolysis to see if you can chop off a disordered region
- consider the
gt;
> --
> *From:* gauri misra
> *To:* CCP4BB@JISCMAIL.AC.UK
> *Sent:* Wed, 23 March, 2011 11:11:55 PM
> *Subject:* [ccp4bb] protein aggregation
>
> Hi,
> What are the different methods to prevent protein aggregation while
> concentrating s
11:11:55 PM
Subject: [ccp4bb] protein aggregation
Hi,
What are the different methods to prevent protein aggregation while
concentrating so as to increase the concentration of the protein?
I have some idea of adding EDTA and charged amino acids like L-Arg and L-Glu.
I would appreciate if the readers
- try limited proteolysis to see if you can chop off a disordered region
- consider the fact that, although it purifies nicely, your protein may not be
well-folded
do you have a biochemical activity test?
Mark J van Raaij
Laboratorio M-4
Dpto de Estructura de Macromoleculas
Centro Nacional de Bio
Try different detergents.
Try 10% or more glycerol.
Try adding ligands (if present/known).
Try varying ionic strength and/or pH.
Try giving more specifics so people on the board may be able to help you better.
Bert
On 3/23/11 1:51 PM, "gauri misra" wrote:
The protein purifies nicely there is
The protein purifies nicely there is no problem in that. Just at the last
step when it is concentrated it starts precipitating beyond a concentration
of 1mg/ml.
Already the purification buffers have the detergent.
On Wed, Mar 23, 2011 at 1:44 PM, Kornelius Zeth <
kornelius.z...@tuebingen.mpg.de> w
Hi,
What are the different methods to prevent protein aggregation while
concentrating so as to increase the concentration of the protein?
I have some idea of adding EDTA and charged amino acids like L-Arg and
L-Glu.
I would appreciate if the readers share their experiences.
Thanks!
Gauri
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