I used the technique in a recent publication for resolving high molecular weight protein complexes in human plasma. (The paper is here: doi: 10.1074/jbc.RA117.001078) In my experience limiting sample diffusion is key, since agarose gels have a much larger pore size and tend to be much thicker. Run-time, voltage, and gel thickness and temperature are all are important variables there. In my hands performing the run with the gel box immersed in ice (to limit heating) with voltage at 5-7 V/cm was ideal. Pouring the thinnest gel possible that was still able to accommodate my sample volume also helped to limit diffusion. As a side-note I have no evidence that the complexes I was detecting in human plasma were amyloids, so things may need to be optimized in your system. It is possible that massive amyloids can be run much slower/longer and still resolved well, since many of the published protocols on this use relatively slow electrophoresis runs.

Hope this helps.
Bill
10/16/2018 12:02:58 PM Reply