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SDS-PAGE/Immunoblot Detection of Aβ Multimers in Human Cortical Tissue Homogenates using Antigen-Epitope Retrieval
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Chapters
- 00:00Title
- 00:47Introduction
- 01:20Preparation of Clarified Tissue Homogenates
- 02:43SDS-PAGE Sample Preparation and Electrophoresis
- 04:12Transferring Proteins from Gels to Membranes
- 05:35Antigen-Epitope Retrieval and Immunoblotting
- 08:13Aβ Multimers by SDS-Page Analysis Using the 6E10 Antibody
- 09:53Conclusion
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.
Tags
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
