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

Labeling DNA Probes03:31

Labeling DNA Probes

8.3K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
8.3K

You might also read

Related Articles

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

Sort by
Same author

A new multi-purpose FRET fluorescent probe for the simultaneous detection of proteases.

RSC advances·2025
Same author

Novel Peptide-Based Fluorescent Probe for Simultaneous Sensing of Chymotrypsin and Hydrogen Peroxide.

ACS omega·2024
Same author

Drug-like Inhibitors of DC-SIGN Based on a Quinolone Scaffold.

ACS medicinal chemistry letters·2022
Same author

Amino-BODIPY as the ratiometric fluorescent sensor for monitoring drug release or "power supply" selector for molecular electronics.

RSC advances·2022
Same author

Polymer-supported synthesis of <i>N</i>-substituted anthranilates as the building blocks for preparation of <i>N</i>-arylated 3-hydroxyquinolin-4(1<i>H</i>)-ones.

RSC advances·2022
Same author

AminoBODIPY Conjugates for Targeted Drug Delivery Systems and Real-Time Monitoring of Drug Release.

Molecular pharmaceutics·2021

Related Experiment Video

Updated: Aug 23, 2025

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
19:23

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Published on: January 16, 2019

9.3K

Triple-FRET multi-purpose fluorescent probe for three-protease detection.

David Milićević1, Jan Hlaváč1

  • 1Department of Organic Chemistry, Faculty of Science, Palacký University Olomouc 17. Listopadu 12 771 46 Olomouc Czech Republic jan.hlavac@upol.cz.

RSC Advances
|November 2, 2022
PubMed
Summary

A novel peptide probe enables sensitive detection of trypsin, chymotrypsin, and caspase-8 proteases. This method allows for single-protease screening but has limitations for simultaneous detection of multiple proteases.

More Related Videos

Use of the Protease Fluorescent Detection Kit to Determine Protease Activity
09:00

Use of the Protease Fluorescent Detection Kit to Determine Protease Activity

Published on: August 4, 2009

28.4K
Detection of Protease Activity by Fluorescent Peptide Zymography
09:56

Detection of Protease Activity by Fluorescent Peptide Zymography

Published on: January 20, 2019

12.9K

Related Experiment Videos

Last Updated: Aug 23, 2025

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
19:23

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Published on: January 16, 2019

9.3K
Use of the Protease Fluorescent Detection Kit to Determine Protease Activity
09:00

Use of the Protease Fluorescent Detection Kit to Determine Protease Activity

Published on: August 4, 2009

28.4K
Detection of Protease Activity by Fluorescent Peptide Zymography
09:56

Detection of Protease Activity by Fluorescent Peptide Zymography

Published on: January 20, 2019

12.9K

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Protease activity is crucial in various biological processes and disease states.
  • Accurate and sensitive detection of specific proteases is essential for research and diagnostics.
  • Existing methods for protease screening can be complex or lack specificity.

Purpose of the Study:

  • To develop a robust and reliable methodology for screening three specific proteases in a single-enzyme mode.
  • To create a multi-purpose peptide probe capable of detecting trypsin, chymotrypsin, and caspase-8.
  • To evaluate the feasibility of simultaneous two-protease screening using the developed probe.

Main Methods:

  • Development and verification of a novel methodology using a multi-purpose peptide probe.
  • The probe features three selectively cleavable sites and four fluorophores.
  • Application of a triple- Förster Resonance Energy Transfer (FRET)-based single-excitation quadruple-emission concept for sensing.

Main Results:

  • Achieved unambiguous sensing of trypsin, chymotrypsin, and caspase-8 at low detectable concentrations.
  • Demonstrated successful single-protease screening with high sensitivity.
  • Found simultaneous two-protease screening to be only partially feasible due to unselective chymotrypsin cleavage.

Conclusions:

  • The developed methodology provides a sensitive and reliable tool for single-protease screening.
  • The multi-purpose peptide probe and FRET-based concept are effective for detecting specific proteases.
  • Further optimization is needed to overcome limitations in simultaneous multi-protease detection.