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
