SDS-PAGE/Immunoblot Detection of A&beta; Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval

Rebecca F. Rosen; Yasushi Tomidokoro; Jorge A. Ghiso; Lary C. Walker

doi:10.3791/1916

Journal

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Neuroscience

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SDS-PAGE/Immunoblot Detection of Aβ Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

SDS-PAGE/Immunoblot Detection of Aβ Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval

23,992 Views

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10:48 min

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April 23, 2010

DOI:

10.3791/1916-v

DOI:

10.3791/1916-v

•

10:48 min

April 23, 2010

•

5 Views

Rebecca F. Rosen¹, Yasushi Tomidokoro², Jorge A. Ghiso³, Lary C. Walker^1,4

¹Yerkes National Primate Research Center,Emory University, ²Department of Neurology, Institute of Clinical Medicine,Tsukuba University, ³Department of Pathology,New York University School of Medicine, ⁴Department of Neurology,Emory University

Summary

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.