electron microscopy of filament formation
10mM Tris-Cl, 160 mM NaCl
Protofilaments are generated by overnight dialysis against a Low Salt buffer. In brief, protein at ∼500 μg/mL in 8M urea was dialyzed overnight against intermediate filament assembly buffer (10 mM Tris‐Cl, pH 7.5 and 160 mM NaCl) and approximately 10 μL of the sample was removed, placed on formvar‐ and carbon‐coated grids then stained with 1% uranyl acetate. Grids were imaged with a Phillips CM‐120 electron microscope operated at 80 kV acceleration voltage. reference (https://doi.org/10.1002/pro.2182).