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Related Concept Videos

Tagging and Fusion Proteins01:24

Tagging and Fusion Proteins

Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...

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Related Experiment Video

Updated: May 15, 2026

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

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An approach for triplex-isobaric peptide termini labeling (triplex-IPTL).

Christian J Koehler1, Magnus Ø Arntzen, Gustavo Antonio de Souza

  • 1The Biotechnology Centre of Oslo, University of Oslo, P.O. Box 1125 Blindern, 0317 Oslo, Norway.

Analytical Chemistry
|January 16, 2013
PubMed
Summary

We developed triplex Isobaric Peptide Termini Labeling (IPTL) for simultaneous quantification of three biological states. This method, combined with the IsobariQ tool, enables robust protein profiling and analysis of cellular responses.

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Automated Sample Multiplexing by using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)
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Published on: December 18, 2020

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
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Published on: July 1, 2014

Related Experiment Videos

Last Updated: May 15, 2026

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

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

Area of Science:

  • Proteomics
  • Chemical Biology
  • Mass Spectrometry

Background:

  • Isobaric peptide termini labeling (IPTL) enables quantitative proteomics by labeling peptide termini with isotopic tags.
  • Current methods may lack the multiplexing capacity for analyzing multiple conditions simultaneously.

Purpose of the Study:

  • To introduce triplex-IPTL, a novel chemical labeling strategy for simultaneous quantification of three states in a single mass spectrometry run.
  • To develop and validate the IsobariQ software for analyzing triplex-IPTL data.

Main Methods:

  • Triplex-IPTL involves N-terminal and lysine dimethylation using stable isotopes of formaldehyde and cyanoborohydride.
  • Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for analysis.
  • The IsobariQ tool was enhanced with multidimensional VSN normalization, statistical inference, and visualization for data analysis.

Main Results:

  • Triplex-IPTL successfully generated three distinct isotopic fragment ion series, enabling quantitative peptide ratio determination.
  • The IsobariQ tool effectively processed triplex-IPTL data, revealing protein ratio profiles.
  • Temporal profiling of HeLa cells treated with STLC identified protein clusters associated with mitotic arrest and cell death.

Conclusions:

  • Triplex-IPTL offers a powerful multiplexing strategy for quantitative proteomics.
  • The integrated IsobariQ software facilitates comprehensive analysis of complex proteomic data.
  • This approach is valuable for studying cellular responses and biological pathway dynamics.