Xenbase Image ID: 126570

Figure 1. Western blot analysis of Aβ1−42 oligomer preparations and assay of potency to evoke Ca2+ influx in Xenopus oocytes. (A and B) Western blots showing aggregation profiles of Aβ1–42 at 30 min after initially dissolving Aβ peptide monomer (A) and after 48 h of incubation (B). 4 µg of samples was separated using gel electrophoresis on a 4–20% Tris-HCl gel. The membranes were probed separately with a monoclonal antibody, 6E10, which is sequence specific to Aβ, and with OC antibody, which recognizes a generic epitope associated with the fibrillar amyloid conformation independent of peptide sequence. Black lines indicate that intervening lanes have been spliced out. (C and D) Bioassay of the potency of Aβ1–42 oligomer preparations to induce membrane permeability to Ca2+. The traces in C show representative global Ca2+ fluorescence signals (indicated as F; top) and corresponding voltage-clamped membrane currents (indicated as I; bottom) evoked by 30-s voltage steps from 0 to −100 mV delivered at various times (indicated in minutes) before and after pipette application of Aβ1−42 oligomers to a nonpeeled oocyte loaded with fluo-4 dextran. (D) Measurements of inward currents (without leak subtraction) evoked by voltage steps from 0 to −100 mV taken at 5-min intervals after application of Aβ1−42 (final bath concentration of 1 µg/ml). Data show means ± 1 SEM from four oocytes using two different Aβ1−42 oligomer preparations. Image published in: Demuro A et al. (2011) © 2011 Demuro et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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