off topic: Another protein forming gel

From: Obayed Ullah
Date: 21 September 2011 04:57


Hi all

I am working with a human protein (not membrane protein) which is forming gel at higher concentration. I am not able to concentrate the protein even more than 7 mg/ml. It become like jelly which is hard to pipette out.

I have gone through all the suggestions which you guys proposed very recently for another member. what i mean that i already have tried different temperature, pH, salt concentration, reducing agents (DTT, TCEF, in different concentration), stability analysis. But unfortunately nothing is working.

One think i want to try to add some detergent to it. If you guys have any experience about it then please suggest the name and/or kind of detergent for the protein. Any other suggestion also appreciated which might help.

cheers

 
M. Obayed Ullah

----------
From: Tim Gruene 

Dear Obayed,

it is not clear to me whether with 'nothing is working' you mean to
concentrate the solution even further or whether you mean that you do
not get crystals from 7mg/ml.

If you refer to the former, I would proceed with crystallisation trials
at 7mg/ml.

Tim

- --
- --
Dr Tim Gruene
Institut fuer anorganische Chemie
Tammannstr. 4
D-37077 Goettingen



----------
From: Obayed Ullah

Hi Tim

Thanks for your mail. Both are true. I have not got crystals yet. I have tired different truncated constructs as well as concentration for crystallization. Even i set up the tray at 2 mg/ml but still proteins are not mixing well with the drop solutions rather sitting on the middle of the drop. I dont know what to do with that one. Please suggest me.