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.
