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SDS-PAGE/Immunoblot Detection of Aβ Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval
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April 23, 2010
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We describe a technique for the preparation of clarified human cortical homogenates, protein separation by SDS-PAGE, antigen retrieval and immunoblotting with an antibody to the Aβ peptide. Using this protocol, we consistently detect monomeric and multimeric Aβ in cortical tissue from humans with Alzheimer's pathology.
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SDS-PAGE/ImmunoblotAβ MultimersAntigen-epitope RetrievalAnomalous FoldingPolymerizationβ-amyloid PeptideNeurodegenerative CascadeAlzheimer’s Disease PathogenesisSelf-aggregationAmyloid FibrilsCerebral Senile PlaquesLow Molecular WeightSoluble Aβ MultimersToxic MultimersProteopathiesSDS-polyacrylamide Gel ElectrophoresisImmunoblottingAβ-specific AntibodiesHighly Concentrated Cortical HomogenatesSmall Pore-size Nitrocellulose MembranesClarified Human Cortical Homogenates
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Rosen, R. F., Tomidokoro, Y., Ghiso, J. A., Walker, L. C. SDS-PAGE/Immunoblot Detection of Aβ Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval. J. Vis. Exp. (38), e1916, doi:10.3791/1916 (2010).
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