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Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting
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JoVE Journal Neuroscience
Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting

Consensus Brain-derived Protein, Extraction Protocol for the Study of Human and Murine Brain Proteome Using Both 2D-DIGE and Mini 2DE Immunoblotting

DOI:
10.3791/51339-v

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10:51 min

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April 10, 2014

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, , , , , , , , , , , , , , ,
1Team Alzheimer & Tauopathies, Jean-Pierre Aubert Research Centre,Inserm UMR 837, 2EA 4308-Department of Reproductive Biology-Spermiology-CECOS,CHRU-Lille, 3EA2686-Laboratorie d’Immunologie,Faculté de Médecine – Pôle Recherche, 4Department of Neurology,CHRU-Lille

Chapters

  • 00:05Title
  • 01:08Extracting Total Protein
  • 03:07Quality Control and Dye Conjugation
  • 05:28Fluorescence Difference Gel Electrophoresis (2D-DIGE)
  • 07:36Mini-2DE for Quantitative Analysis
  • 08:53Proteomics of Human and Mouse Tissue with Alzheimer Disease
  • 10:26Conclusion

Summary

Automatic Translation

Automatic Translation

A common protein extraction protocol using urea/thiourea/SDS buffer for human and mice brain tissue allows indentification of proteins by 2D-DIGE and their subsequent characterization by mini 2DE immunoblotting. This method enables one to obtain more reproducible and reliable results from human biopsies and experimental models.

Tags

Keywords: Brain Proteome2D-DIGEMini 2DE ImmunoblottingSample PreparationHuman Post-mortemProtein AggregatesAmyloid-betaAlpha-synucleinNeurodegenerative DiseasesBiomarkersTherapeutic Targets

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