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Related Concept Videos

Mass Analyzers: Common Types01:19

Mass Analyzers: Common Types

The quadrupole mass analyzer consists of four cylindrical metal rods arranged in a diamond carrying a DC voltage and a radio-frequency AC voltage. The motion of ions through the quadrupole depends on the field strength, causing only ions of a certain m/z to resonate successfully and strike the detector at a given field strength. Though the transmission rate for these analyzers is high, the exact elemental composition of the sample is not determined because of low resolution; however, they are...

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Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling
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Published on: November 17, 2019

High-throughput middle-down analysis using an orbitrap.

Joe Cannon1, Karen Lohnes, Colin Wynne

  • 1Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland 20274, USA.

Journal of Proteome Research
|June 19, 2010
PubMed
Summary

This study presents an automated middle-down proteomic analysis workflow for large peptides using capillary LC-LTQ-orbitrap. The novel method enhances mass accuracy for ribosomal proteome analysis in human cancer cells.

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Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)
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Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Published on: May 20, 2013

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Middle-down proteomics offers advantages for analyzing large peptides.
  • Orbitrap mass spectrometry provides high resolution and mass accuracy.
  • Automated workflows are crucial for efficient proteomic analysis.

Purpose of the Study:

  • To develop and demonstrate an automated middle-down proteomic analysis workflow.
  • To utilize high-resolution mass-accurate product ion measurements in Orbitrap.
  • To analyze the ribosomal proteome of human MCF7 cancer cells.

Main Methods:

  • Capillary Liquid Chromatography-Linear Trap Quadrupole-Orbitrap (LC-LTQ-Orbitrap) system.
  • Automated middle-down analysis of proteolytic peptides (3000-10,000 Da).
  • Asp-selective chemical cleavage and ProSightPC 2.0 software for data searching.

Main Results:

  • Successful application of the LC-LTQ-orbitrap system for automated middle-down analysis.
  • High-resolution, mass-accurate precursor and product ion measurements achieved.
  • Demonstrated analysis of the highly basic ribosomal proteome from human MCF7 cancer cells.

Conclusions:

  • The novel workflow enables efficient and accurate middle-down proteomic analysis.
  • High-resolution mass measurements are key for analyzing complex proteomes.
  • This approach is valuable for studying cellular proteomes, including cancer research.