Triton _X-114

MJ Duggan sgex400 at sghms.lon.ac.uk
Thu Nov 25 12:24:57 EST 1993


Dear Netters,

Has anybody out there used Triton-X-114 for the phase separation of 
membrane proteins?

I am trying this following the method of Bordier (1981) J. Biol. Chem.
_265_, 1604-1607. In this he ends up by spinning the T-X-114 phase down 
through a sucrose cushion. The problem that I have is that my T-X-114
phase floats above the buffer! (I am only washing the T-X-114 at present).
The only difference that I can see between what I am doing and what is
in the paper is that I am using PBS whereas he used Tris buffered saline
(150 mM NaCl).

Can anybody help. Am I doing anything wrong or is the paper wrong or
misreadable? Any help would be welcome, I will post a summary if there is any 
interest.

Thanks in advance,

Michael Duggan



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