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Peptide Identification Using Tandem Mass Spectrometry01:33

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...

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Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
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Solid phase isobaric mass tag reagent for simultaneous protein identification and assay.

Anna Napoli1, Constantinos M Athanassopoulos, Petros Moschidis

  • 1Dipartimento di Chimica, UniversitĂ  della Calabria, Arcavacata di Rende, Italy. amc.napoli@unical.it

Analytical Chemistry
|June 10, 2010
PubMed
Summary

Solid phase isobaric mass tagging (SPIMT) enables simultaneous protein quantitation and identification. This method uses CID-based differentiation of tagged peptides for accurate, low-cost, two-plex protein analysis.

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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Accurate protein quantitation is crucial in biological research.
  • Existing methods for multiplexed protein quantitation can be complex or costly.

Purpose of the Study:

  • To introduce a novel Solid Phase Isobaric Mass Tagging (SPIMT) approach.
  • To enable simultaneous protein quantitation and identification using a single method.

Main Methods:

  • SPIMT utilizes unlabeled mass-tagging reagents linked to N-terminal peptides.
  • Quantitation is achieved through CID-based differentiation of regioisomeric tagged peptides.
  • Peptide identification is facilitated by direct submission of MS/MS data to MASCOT.

Main Results:

  • SPIMT allows for the quantitation of tagged peptides via unique reporter ion masses (m/z 168.2 and 182.2 Da).
  • The method demonstrates excellent quantitation linearity over a 5-fold concentration range.
  • Sequencing information is preserved during the quantitation process.

Conclusions:

  • SPIMT offers a simple, low-cost alternative for two-plex protein quantitation.
  • The SPIMT strategy is adaptable, with potential for tuning signal mass windows.