Two Phase Separation

M. Johan Broekman hanbroekman at csi.com
Wed Jan 5 21:23:53 EST 2000


"A. J. Bowling" wrote:
> 
> hello all,
> 
> i am trying to obtain some plasma membrane using an aqueous two phase system
> containing dextran and peg.  everything is fine until i put my microsomal
> fraction onto the two phase mixture and mix them.  no amount of centrifugation
> or sitting at 4 deg. will cause the two phases to separate again.  does anyone
> know what is going on here?
> 
> thanks for taking the time to read this.  believe me, any help will be greatly
> appreciated.
> 
> andy b

Any 2-phase system will be perturbed by "detergent".  The microsomal
prep may be acting as such (lots of lipid).  Also, did you take into
account the quantity of liquid introduced by the microsomes?  What
happens when you mix the same volume of buffer (as used to resuspend the
microsomes) into the 2-phase system?
-Han




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