I am trying to perform a native gel electrophoresis assay on my protein. I made a 4-12% gradient gel just like I would make a denaturing gel, but omitted SDS. I ran the gel with a Tris-glycine running buffer and prepared my samples with 2x sample buffer containing Tris-HCl pH 6.8, glycerol, and bromophenol blue.
I did not observe any dimerization of my protein as I expected. I am aware that this simply means that my protein does not form dimers. However, I am afraid that I might have done something to denature my protein.
I want to say that making Tris-glycine gels would be better than the other gels I made (as I have found these more often used in the literature) but I cannot find a recipe for them.
Does anyone have a recipe or can provide insight on how to go about performing this assay?
Take a look at this paper. They made the running buffers with 192 mM glycine and 50 mM Tris without SDS for native PAGE.
http://onlinelibrary.wiley.com/doi/10.1002/jssc.201100315/abstract
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