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

You might also read

Related Articles

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

Sort by
Same author

Proton Transfer Charge Reduction Enables Isobaric Labeling-Based Proteoform Quantification of Overlapping Signals in Top-Down Mass Spectrometry.

Journal of the American Society for Mass Spectrometry·2026
Same author

ADAM10-Mediated Proteolytic Remodelling of Signalling and Adhesion Proteins on Brain Cell-Derived Small Extracellular Vesicles.

Journal of extracellular biology·2026
Same author

H<sub>2</sub>-dependent modulation of tetrahydromethanopterin S-methyltransferase (Mtr complex) activity by the small protein MtrR in Methanosarcina mazei.

The FEBS journal·2026
Same author

CoMPaseD: advanced planning of proteomic experiments aiming to identify small proteins.

microLife·2026
Same author

An ancient lysozyme in placozoans participates in acidic extracellular digestion.

Communications biology·2026
Same author

Meprin β elevates hippocampal soluble Aβ in the APP/V717I mouse model.

Experimental neurology·2025
Same journal

Isolation of Mesenchymal Stem Cell-Derived Extracellular Vesicles.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Modeling Melanoma Immune Surveillance by CAR-T Cells in Human Skin Organoids.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Stepwise Optimization of a Matrigel-Based In Vitro Angiogenesis Assay for Reproducible and Quantifiable 2D-Tube Formation Using HUVECs.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Quantifying Mechanical Properties of Fresh Ovarian Tissue with Optical Brillouin Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

3D Chromatin Architecture During Early Development: New Methods and New Findings.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Metabolic Plasticity in Embryogenesis Throughout the Lens of NAD<sup></sup>.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: Mar 6, 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

7.5K

Multiplexed Protease Specificity Profiling Using Isobaric Labeling.

Joanna Tucher1, Andreas Tholey2

  • 1AG Systematic Proteome Research & Bioanalytics, Institute for Experimental Medicine, Christian-Albrechts-Universität zu Kiel, Campus UKSH-Kiel, Haus 1, Niemannsweg 11, 24105, Kiel, Germany. j.tucher@iem.uni-kiel.de.

Methods in Molecular Biology (Clifton, N.J.)
|March 19, 2017
PubMed
Summary
This summary is machine-generated.

This study details a quantitative proteomic identification of protease cleavage sites (Q-PICS) method. It enables precise comparison of protease activity and cleavage specificity under various conditions using tandem mass tags (TMT).

Keywords:
Cleavage site specificityHeat mapIsobaric labelingLC-MSProteaseProteome-derived peptide library

More Related Videos

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

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

Published on: November 15, 2017

12.8K
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

6.1K

Related Experiment Videos

Last Updated: Mar 6, 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

7.5K
Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

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

Published on: November 15, 2017

12.8K
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

6.1K

Area of Science:

  • Proteomics
  • Enzymology
  • Biochemistry

Background:

  • Determining protease cleavage site specificity is crucial in degradomics.
  • Proteomic identification of protease cleavage sites (PICS) using peptide libraries and LC-MS is a common approach.
  • Quantitative analysis allows for comparative studies of protease activity.

Purpose of the Study:

  • To provide a detailed protocol for a quantitative proteomic identification of protease cleavage sites (Q-PICS) experiment.
  • To enable relative quantification of proteolytic events using isobaric labeling.
  • To facilitate comparison of protease cleavage specificity and activity under varying experimental conditions.

Main Methods:

  • Utilizes proteome-derived peptide libraries.
  • Employs liquid chromatography-mass spectrometry (LC-MS).
  • Incorporates isobaric labeling, such as tandem mass tags (TMT), for quantitative analysis.
  • Leverages multiplexing for parallel analysis of replicates and internal controls.

Main Results:

  • The Q-PICS method allows for relative quantification of proteolytic events.
  • Enables comparison of protease cleavage site specificity and activity across different conditions (e.g., buffer, pH, temperature, inhibitors).
  • Multiplexing enhances efficiency by analyzing replicates and controls in parallel, reducing instrument time and workload.

Conclusions:

  • The Q-PICS protocol offers a robust method for detailed analysis of protease function.
  • This quantitative approach is valuable for understanding protease behavior in complex biological systems.
  • The method supports comparative degradomics studies and drug discovery efforts.