Refining Western Blot Procedures: A Modified Approach for the Reliable Detection of Amyloid-Beta Oligomers (AβOs) in Synthetic, Mice, and Human Tissues For Alzheimer's Research
Session Number
2
Advisor(s)
Dr. Raquel de Campos, PhD; Kirsten L Viola; Dr. William Klein, PhD. Klein Lab, Northwestern University
Location
A121
Discipline
Medical and Health Sciences
Start Date
15-4-2026 11:10 AM
End Date
15-4-2026 11:55 AM
Abstract
Neurotoxic amyloid-beta oligomers (AβOs) accumulate in Alzheimer’s Disease patients, driving cognitive decline and dementia. Determining their size and distinguishing them from benign or beneficial oligomers via Western Blots (WB) could help in the development of targeted therapeutics. However, this analysis is particularly challenging for AβO aggregates, given their inconsistent accumulation and tendency to dissociate upon exposure to detergents such as sodium dodecyl sulfate (SDS). Such inconsistencies have led researchers to avoid WB for both one-dimensional (1D-) and two-dimensional gel electrophoresis (2D-PAGE) on AβOs. This study aims to re-establish WB as an effective technique for studying AβOs by varying the initial SDS concentration in the sample and the running buffer, identifying optimal conditions where dissociation does not occur. We also attempt to establish 2D-PAGE as a technique for studying whole-proteomic distribution across both size and pH in AβOs. Our preliminary results indicated new ideal SDS conditions for the running and sample buffers, revealing proteins around the 37 kDa mark that weren’t visible in classic SDS-PAGE. Similarly, 2D-PAGE with SDS can separate synthetic AβOs, wild-type/5xFAD-transgenic mouse samples, and human brain samples with very high resolution across both pH and molecular weight, holding great potential to provide novel insights into AβO properties.
Refining Western Blot Procedures: A Modified Approach for the Reliable Detection of Amyloid-Beta Oligomers (AβOs) in Synthetic, Mice, and Human Tissues For Alzheimer's Research
A121
Neurotoxic amyloid-beta oligomers (AβOs) accumulate in Alzheimer’s Disease patients, driving cognitive decline and dementia. Determining their size and distinguishing them from benign or beneficial oligomers via Western Blots (WB) could help in the development of targeted therapeutics. However, this analysis is particularly challenging for AβO aggregates, given their inconsistent accumulation and tendency to dissociate upon exposure to detergents such as sodium dodecyl sulfate (SDS). Such inconsistencies have led researchers to avoid WB for both one-dimensional (1D-) and two-dimensional gel electrophoresis (2D-PAGE) on AβOs. This study aims to re-establish WB as an effective technique for studying AβOs by varying the initial SDS concentration in the sample and the running buffer, identifying optimal conditions where dissociation does not occur. We also attempt to establish 2D-PAGE as a technique for studying whole-proteomic distribution across both size and pH in AβOs. Our preliminary results indicated new ideal SDS conditions for the running and sample buffers, revealing proteins around the 37 kDa mark that weren’t visible in classic SDS-PAGE. Similarly, 2D-PAGE with SDS can separate synthetic AβOs, wild-type/5xFAD-transgenic mouse samples, and human brain samples with very high resolution across both pH and molecular weight, holding great potential to provide novel insights into AβO properties.