Notably, the compaction of filaments over time is much higher in the "Tris-buffer" (about 17 to 10 nm) as compared to the "phosphate-buffer" assembly mode (about 12 to 9.5 nm).
Minute amounts of dimers or higher order species may be present at high concentrations of CLR01 in some cases in PBS buffer (fibril assembly and cell culture buffer), but are absent from the HEPES buffer (fibril remodeling buffer).
Vimentin was assembled in phosphate assembly buffer at 0.2 mg/ml for about 25 min. Then, the sample was diluted 40 to 60 fold in the same buffer and applied to the surface.
Minute amounts of dimers or higher order species may be present at high concentrations of CLR01 in some cases in PBS (fibril assembly buffer) but are absent from the HEPES buffer (fibril remodeling buffer).
After blowing HFIP from the Aβ1 42 stock aliquot, the dried peptide (50 μM) was suspended in desired volume of assembly buffer (100 μl of 50-mM Tris HCl buffer, pH 7.4, containing 100 mM NaCl and 0.01% sodium azide) with or without asiaticoside (final concentrations of asiaticoside were 0, 5, 10, and 20 μM).
Labeled wild-type and mutant yeast αβ-tubulin were polymerized in the presence of 3 5 µM epothilone-B in assembly buffer (see above) with 1 mM GTP or in the presence of 500 µM GTPγS in assembly buffer (see above).
Optimizing the assembly buffer exchange rate, and the rapid introduction of calcium ions to the assembly mix, facilitated the propitious assembly of VLPs.
Thus, we expected to observe only individual microtubules in a commonly used microtubule assembly buffer containing fluorescently labeled tubulin and 20 µM Taxol (as in Materials and methods).
The sample was either diluted or quenched by addition of an equal volume of 0.2% glutaraldehyde in the corresponding assembly buffer.
Assembled filaments were deposited onto glass modified to be hydrophilic or freshly cleaved mica in 2 mM sodium phosphate, pH 7.5, 100 mM KCl [13], [25] (phosphate assembly buffer).
