Figure 1.
In vitro microdialysis sampling of HMW Aβ oligomers by using a large-pore-sized (1000-kDa-MWCO) membrane probe. A–C) An Aβ oligomer mixture was prepared by incubating synthetic human Aβ1–42 (0.1 mg/ml) for 3 h at 37°C. Concentrations of Aβ in preincubation and postincubation samples were measured by 82E1-82E1 ELISA for Aβ oligomers (A) and BAN50-BC05 ELISA for Aβ1–42 (B). Samples were treated with and without guanidine HCl before ELISA measurement (n=3/group). **P < 0.01. C) SEC of the preincubation and postincubation Aβ1–42 solutions was performed on a Superdex75 column. Aβ levels in each SEC-separated sample were measured by ELISA (BAN50-BC05). The preincubation sample was treated with guanidine HCl to completely dissociate Aβ oligomers. D, E) In vitro microdialysis with small- and large-pore-sized (35- and 1000-kDa-MWCO) membranes from Aβ oligomer mixtures. D) Microdialysis samples were collected from the same Aβ oligomer mixture solution by using 35- and 1000-kDa-MWCO membranes at a flow rate of 1.0 μl/min. E) Microdialysis samples and the original solution (Aβ oligomer mixtures) were separated by SEC, followed by Aβ quantification of each SEC fraction (BAN50-BC05 ELISA). Mo, monomer; MWM, molecular weight marker.