A protein is expected to form multimers in solution. How can I establish this fact? What experiments should I perform apart from Size exclusion chromatography and native PAGE?
there are several methods to ensure dimer formation of protein.
1.Do gel permeation chromatography and compare molecular weight ( Mol wt calculated from gene sequence).
2. Perform cross linking( eg. glutaraldehyde cross linking) followed the SDS PAGE.
3. Through Native page
and so on..
Thank you very much Adam for the suggestion. Also, I was wondering if surface plasmon resonance could be used to establish dimer formation? Any advise on the matter would be helpful.
Thanks Gopal Jee Gopal. I could not quite understand the cross linking part but I will look it up. Many thanks for your guidance and help in this regard.
I can't say for sure that SPR isn't suitable for this purpose, but I think it would be a complicated method to develop, and the data would be difficult to interpret.
Size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) is an excellent method for measuring the oligomeric state of a protein, if you know the monomer mass and the equipment is available.
As for using crosslinking to establish the oligomeric state of a protein, please see the attached paper:
Thank you very much Adam Shapiro sir and Kou Hayakawa sir. I will go through all the information provided by you and proceed accordingly. Many thanks once again.
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