I work with a natural adhesive (footprint) extracted from a marine organism onto the glass slide or mica. It is very cohesive sample and so far, we have detected amyloid fibrils in the footprint (in combination with many other proteins and they have not been completely identified yet). My goal is to make high resolution images of amyloid fibrils and to do that I need to selectively wash away other components/proteins in the footprint. I probably should try some detergent but can anyone suggest which detergent to use and concentration?
There are several examples of cleaning solutions you could use that are commonly employed in protocols that extract amyloids from natural sources, mainly brain tissue, such as prions and other amyloids associated with neurodegenerative disorders. A detergent I have used in the past for a similar task is N-lauroylsarcosine (Sarkosyl). I refer you to a PNAS paper by Prusiner that you could use as starting point for developing a suitable washing method (http://www.pnas.org/content/104/52/20914.full). Good luck with the experiments.
I agree with both Carlos and Kai in the use of N-lauroylsarcosine in unison with ultracentrifugation please refer to:-
(2004) Michael J. Stack Western Immunoblotting Techniques for the Study of. Transmissible Spongiform Encephalopathies. ... Biosciences and Medicine – Techniques in Prion Research. ... Birkhäuser Verlag, Berlin. Chapter 8, 97- 116
This gives flow charts, g forces and the correct concentrations of Sarkosyl to clean up scrapie-associated fibrils prior to immunoblotting.
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