Dear Careina,
Could you tell what reason you have to believe that your protein is
oligomerising? If your protein is forming oligomers, you should be able
to detect them using crosslinking or Native PAGE.
Do you have access to an analytical ultracentrifuge?
cheers
Ganesh
Le 14/11/12 08:56, Careina Edgooms a écrit :
Anyone have any advice on how to detect whether my protein is forming
oligomers? It is monomeric in the native state but I have reason to
believe that it may be oligomerising in mild concentrations of urea
(intermediate state).
I have tried cross linking and BN PAGE and they are inconclusive.
SE-HPLC does not work because at the concentrations required to
produce a signal, this intermediate species aggregates.
I do not have access to an ultracentrifuge. The dynamic light
scattering equipment that is available to me is really a poor
instrument which will not be sensitive enough to pick up changes in
size (we are looking at about 5nm for the monomer and anything from
8nm for the oligomer). The only other option I can think of is SAXS. I
will only be able to use that equipment in the middle of next year.
I'm wondering if there is anything else, any other technique or idea
that I have not thought about that I could try? I really just would
like to show that the stoichiometry of the intermediate species is a
multiple of the native state.
If anyone has any suggestions, that would be great.
Thanks
Careina.
