Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules

Maximilian Wulf; Katalin Barkovits-Boeddinghaus; Paula Sommer; Karin Schork; Martin Eisenacher; Peter Riederer; Manfred Gerlach; Steffen K&#246;sters; Britta Eggers; Katrin Marcus

doi:10.3791/63289

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Protocollo basato sulla microdissezione laser per l'analisi LC-MS/MS del profilo proteomico dei granuli di neuromelanina

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Neurosciences

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Journal JoVE Neurosciences

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules

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Protocollo basato sulla microdissezione laser per l'analisi LC-MS/MS del profilo proteomico dei granuli di neuromelanina

2,001 Views

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07:35 min

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December 16, 2021

DOI:

10.3791/63289-v

DOI:

10.3791/63289-v

•

07:35 min

December 16, 2021

•

32 Views

Maximilian Wulf^1,2, Katalin Barkovits-Boeddinghaus^1,2, Paula Sommer^1,2, Karin Schork^1,2, Martin Eisenacher^1,2, Peter Riederer^3,4, Manfred Gerlach⁵, Steffen Kösters^1,2, Britta Eggers*^1,2, Katrin Marcus*^1,2

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

Summary

Un robusto protocollo è presentato qui per isolare i granuli di neuromelanina dal tessuto umano post-mortem substantia nigra pars compacta tramite microdissezione laser. Questo protocollo rivisto e ottimizzato riduce drasticamente il tempo necessario per la raccolta dei campioni, riduce la quantità di campione richiesta e migliora l'identificazione e la quantificazione delle proteine mediante analisi LC-MS/MS.

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Laser Microdissection Proteomic Profile Neuromelanin Granules Mass Spectrometry Proteomic Analysis Disease Mechanisms Robo Stage Field Of View Analysis Laser Settings Sample Collection Triptic Digestion HPLC-MS Analysis Proteomic Raw Data Analysis

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Protocollo basato sulla microdissezione laser per l'analisi LC-MS/MS del profilo proteomico dei granuli di neuromelanina

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