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Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules
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1Medizinisches Proteom-Center, Medical Faculty,Ruhr-University Bochum, 2Medical Proteome Analysis, Center for Proteindiagnostics (PRODI),Ruhr-University Bochum, 3Center of Mental Health; Clinic and Policlinic for Psychiatry, Psychosomatics and Psychotherapy,University Hospital Wuerzburg, 4Psychiatry Department of Clinical Research,University of Southern Denmark Odense University Hospital, 5Center of Mental Health, Department of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy,University Hospital of Wuerzburg, University of Wuerzburg
Chapters
- 00:05Introduction
- 00:46Laser Microdissection and Pressure Catapulting
- 02:46Sample Preparation for HPLC-MS and Proteomic Raw Data Analysis
- 05:10Statistical Analysis
- 05:50Results: Protein Identification in Neuromelanin Granules (NMGs) and Substantia nigra (SN)
- 07:00Conclusion
A robust protocol is presented here for isolating neuromelanin granules from human post-mortem substantia nigra pars compacta tissue via laser microdissection. This revised and optimized protocol massively minimizes the required time for sample collection, reduces the required sample amount, and enhances the identification and quantification of proteins by LC-MS/MS analysis.
