asymmetric flow field flow fractionation affff system  (Waters Corporation)

Journal: Open Biology

Article Title: Soluble Aβ aggregates can inhibit prion propagation

doi: 10.1098/rsob.170158

Figure Lengend Snippet: Characterization of Aβ species. AFFFF is a chromatography technique in which the separation of the sample is performed in a channel rather than a column, and separation of differently sized particles is achieved with two perpendicular flow directions of elution buffer. In AFFFF, small particles elute first, thus the elution profile is the inverse order of size exclusion chromatography. About 190 µl of each Aβ preparation was injected and eluted in 50 mM ammonium acetate pH 8.5 at 1 ml min −1 . Each AFFFF plot shows absorbance at 275 nm throughout the run (blue line) and molar mass (red dots) across the main UV peak. Only the main UV peak contained enough protein for the molar mass to be accurately calculated—see the molar mass of the buffer-only sample ( d ) as an example. Data are shown for Aβ40 monomer ( a ), ADDLs ( b ), Aβ42 fibrils ( c ) and Opti-MEM only ( d ). The EM image for each sample is also shown. Scale bars: ( a , b , d ) 100 nm, ( c ) 500 nm.

Article Snippet: Aliquots of Aβ preparations (190 μl) were injected onto an Eclipse DualTec AFFFF (Wyatt Technology, Santa Barbara, CA, USA) and eluted with 50 mM ammonium acetate pH 8.5.

Techniques: Chromatography, Size-exclusion Chromatography, Injection