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Just a possibility for salvage of your already set-up drops:

 

You can spike the reservoirs with some highly concentrated precipitant (no
matter what as long as

it sucks more water out of your drop). It does not solve your problem but
maybe you can 

revive a few drops and get more information from your experiment.

I use a syringe with a needle and poke through the tape into the reservoir
and top it off with the

high conc. precip. The tiny hole is easy to re-tape and does not hinder
observation.

 

Best, BR 

 

From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of
Mattiroli,Francesca
Sent: Tuesday, February 17, 2015 5:23 AM
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