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Omfattende arbejdsgang af massespektrometri-baserede Shotgun Proteomics af vævsprøver
JoVE Journal
Biology
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JoVE Journal Biology
Comprehensive Workflow of Mass Spectrometry-based Shotgun Proteomics of Tissue Samples
DOI:

14:51 min

November 13, 2021

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Chapters

  • 00:00Introduction
  • 00:42Tissue Lysate Preparation
  • 01:51Quantification and Quality Check of Tissue Lysates
  • 02:33Enzymatic Digestion of Proteins
  • 03:57Desalting of Digested Peptides
  • 05:16Quantification of Desalted Peptides
  • 05:58Labeling of Digested Peptides using iTRAQ Reagents
  • 08:14Liquid Chromatography Set-up
  • 09:25Mass Spectrometry Set-up
  • 12:30Representative Results
  • 13:45Conclusions

Summary

Automatic Translation

Den beskrevne protokol giver en optimeret kvantitativ proteomics analyse af vævsprøver ved hjælp af to tilgange: etiket-baseret og etiket fri kvantitet. Etiketbaserede tilgange har den fordel, at proteiner kvantificeres mere præcist, mens en etiketfri tilgang er mere omkostningseffektiv og bruges til at analysere hundredvis af prøver af en kohorte.

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