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Quantitative Proteomics Workflow using Multiple Reaction Monitoring Based Detection of Proteins from Human Brain Tissue
JoVE Journal
Biology
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JoVE Journal Biology
Quantitative Proteomics Workflow using Multiple Reaction Monitoring Based Detection of Proteins from Human Brain Tissue
DOI:

11:49 min

August 28, 2021

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Chapters

  • 00:05Introduction
  • 00:49Protein Extraction from Brain Tissue
  • 02:04Protein Quantification and Quality Check
  • 02:32Protein Digestion
  • 03:58Desalting and Peptide Quantification
  • 05:06Transition List Preparation of Finalised Targets
  • 07:30LC Parameters
  • 08:03MS Parameters
  • 08:25Run Sequence and Instrument QC
  • 09:08Method Refinement
  • 10:58Results and Conclusions

Summary

Automatic Translation

The protocol aims to introduce the use of a triple quadrupole mass spectrometer for Multiple Reaction Monitoring (MRM) of proteins from clinical samples. We have provided a systematic workflow starting from sample preparation to data analysis for clinical samples with all the necessary precautions to be taken.

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