Help! Dimerisation exp.

Mr. G. Morley gmorley at hgmp.mrc.ac.uk
Wed Sep 4 10:51:45 EST 1996


Hello all..I was wondering of there was a proceedure
to do the following experiment.....

a) I have a mutant receptor which may be active as a monomer.
The normal receptor is dimeric when active.
b) However when the receptor is precipitated it dimerises In Vitro.
c) Idealy I would like to irreversibly dimerise the receptor
In Vivo, wash away the cross-linking agent,lyse and ppt the receptor.
d) The idea being that any In Vitro dimerisation will be reversible
so that when the protein is ran on a denaturing acrylamide gel
any In vitro dimers will break up and run as monomers and any
true In Vivo dimers will remain as dimers.
Whew! Hope this makes sense :)
I would be obliqued if: a) Anyone could sugest a cross linking
agent which causes irreversible dimerisation of proteins..
b)  And not too toxic..for my cells.
If anyone has a methadolgoy they use for dimerisation experiments
that would be really helpful.
Thanks in advance..
Gary Morley.
gmorley at rpms.ac.uk




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