 |
 |
Set up your trays with N:1 ratio of protein:well sol’n, where N > 1, up to, say 5:1? That will tend to concentrate the protein in the drop above the original
concentration.
JPK
From: CCP4 bulletin board [mailto:[log in to unmask]]
On Behalf Of Mattiroli,Francesca
Sent: Monday, February 16, 2015 11:23 PM
To: [log in to unmask]
Subject: [ccp4bb] how to reduce protein solubility
Hi all,
I am struggling with a protein complex that is too soluble. I have reached about 20 mg/ml but I still observe very little precipitation (clear drops in 90-95% of the tested conditions). The proteins are expressed in insect cells and going to higher concentration
is not easily achievable.
I have tried different buffer conditions (salt concentration and pH) and I am testing temperatures. I am at a loss with what to try next.
Do you think PTMs (phosphorylation, acetylation) might be causing this?
Any input on how to decrease solubility?
Thank you very much in advance,
Francesca