denaturing proteins in gel - (Mar/19/2009 )
I'm interested in determining if my protein is running as a dimer (or higher). I have a peptide antibody that works on denatured protein but when I ran a non-reducing gel, the antibody did not recognize the protein. I would like to be able to denature the protein in the (non-reducing) gel after it has run to determine if the protein is running as an oligomer. Does anyone know how I would go about denaturing the protein in the gel?
you could try soaking in sds and 2-me (or dtt).
or you could try transferring then soaking in sds and 2-me to denature on the membrane.