Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Identification of CMTM6 and CMTM4 as PD-L1 protein regulators.

Nature·2017
Same author

Elucidating crosstalk mechanisms between phosphorylation and O-GlcNAcylation.

Proceedings of the National Academy of Sciences of the United States of America·2017
Same author

Natural products triptolide, celastrol, and withaferin A inhibit the chaperone activity of peroxiredoxin I.

Chemical science·2017
Same author

A community proposal to integrate proteomics activities in ELIXIR.

F1000Research·2017
Same author

3-Hydroxybenzoate 6-Hydroxylase from <i>Rhodococcus jostii</i> RHA1 Contains a Phosphatidylinositol Cofactor.

Frontiers in microbiology·2017
Same author

Erratum: Microtubule minus-end regulation at spindle poles by an ASPM-katanin complex.

Nature cell biology·2017

Related Experiment Video

Updated: Jun 24, 2026

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
11:53

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

Published on: July 1, 2014

Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics.

Paul J Boersema1, Reinout Raijmakers, Simone Lemeer

  • 1Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.

Nature Protocols
|March 21, 2009
PubMed
Summary

Stable isotope dimethyl labeling offers a cost-effective method for precise protein quantification in proteomics. This peptide-level technique is automatable for high-throughput studies, using inexpensive reagents applicable to diverse samples.

More Related Videos

Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
09:06

Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

Published on: May 1, 2017

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
09:24

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

Published on: December 18, 2020

Related Experiment Videos

Last Updated: Jun 24, 2026

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
11:53

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

Published on: July 1, 2014

Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
09:06

Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

Published on: May 1, 2017

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
09:24

Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

Published on: December 18, 2020

Area of Science:

  • Proteomics
  • Biochemistry
  • Analytical Chemistry

Background:

  • Accurate protein expression quantification is crucial in modern proteomics.
  • Differential stable isotope incorporation is a common method for relative protein quantification.
  • Existing methods like SILAC and iTRAQ have limitations in cost or sample applicability.

Purpose of the Study:

  • To present stable isotope dimethyl labeling as a reliable, cost-effective, and versatile proteomics technique.
  • To detail protocols for implementing stable isotope dimethyl labeling across various sample amounts.
  • To highlight the advantages of dimethyl labeling over other quantitative proteomics approaches.

Main Methods:

  • Stable isotope incorporation at the peptide level using dimethyl labeling.
  • Development of in-solution, online, and on-column protocols.
  • Application to sample amounts ranging from sub-micrograms to milligrams.
  • Integration with liquid chromatography-mass spectrometry for analysis.

Main Results:

  • Dimethyl labeling is a cost-effective and undemanding procedure.
  • The labeling process is automatable and suitable for high-throughput proteomics.
  • The method uses inexpensive reagents and is applicable to virtually any sample.
  • Full protocol completion, including sample prep and analysis, takes 1.5-3 days.

Conclusions:

  • Stable isotope dimethyl labeling is a highly advantageous technique for relative protein quantification.
  • Its cost-effectiveness, automation potential, and broad applicability make it ideal for high-throughput proteomics.
  • The described protocols facilitate the implementation of this method across diverse research settings.