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 Experiment Videos

F Rodier1, M Hill, B Arrio

  • 1Laboratoire de Biologie Physico-Chimique, Faculté des Sciences, 91, Orsay, France

FEBS Letters
|December 11, 1970
PubMed
Summary
This summary is machine-generated.

7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-chloride) is a reactive fluorogenic reagent that labels trypsin, primarily at lysine residues. Its use in fluorescence depolarization studies reveals temperature-dependent enzyme relaxation times.

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

[Screening and healthcare for pregnant women with psycho-social vulnerability : A French national study].

Gynecologie, obstetrique, fertilite & senologie·2022
Same author

Polyomavirus Large T Antigen Interact with the DISC and Protect against Fas Induced Apoptosis.

TheScientificWorldJournal·2018
Same author

Accumulation of sucrose in vacuoles isolated from red beet tissue.

Planta·2014
Same author

Mitochondrial DNA damage induces apoptosis in senescent cells.

Cell death & disease·2013
Same author

Chronic antiplatelet therapy and mortality among patients with infective endocarditis.

Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases·2009
Same author

Enzyme-catalyzed gel proteolysis: an anomalous diffusion-controlled mechanism.

Biophysical journal·2003
Same journal

Investigating transcription factor dynamics in health and disease using FRAP.

FEBS letters·2026
Same journal

Regulation of CFTR stability at the plasma membrane-Mechanisms and therapeutic opportunities in cystic fibrosis.

FEBS letters·2026
Same journal

Identification of a Shiga toxin A-derived peptide internalized into Gb3 receptor-bearing cells via interaction with the Shiga toxin B subunit.

FEBS letters·2026
Same journal

The dual role of lectins in cancer-immunotherapy tools and therapeutic targets.

FEBS letters·2026
Same journal

Decoding the dynamic extracellular matrix in cancer-3D models and bioscaffolds rewire the rules of tumor progression.

FEBS letters·2026
Same journal

Extending the classical sequence-structure-function paradigm through protein dynamics and context-dependent behavior.

FEBS letters·2026
See all related articles

Area of Science:

  • Biochemistry
  • Chemical Biology
  • Enzymology

Background:

  • 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-chloride) is a known fluorogenic reagent.
  • It reacts with amino and sulfhydryl groups.

Purpose of the Study:

  • To evaluate NBD-chloride as a labeling reagent for trypsin.
  • To investigate enzyme dynamics using fluorescence depolarization.

Main Methods:

  • Labeling of trypsin with NBD-chloride.
  • Thin-layer chromatography for residue analysis.
  • Fluorescence depolarization measurements.
  • Absorbance spectroscopy at 470 nm.

Main Results:

  • NBD-chloride rapidly formed a stable, fluorescent conjugate with trypsin.

Related Experiment Videos

  • Lysine residues were preferentially labeled.
  • Fluorescence depolarization revealed strong temperature dependence of enzyme relaxation time.
  • Enzyme relaxation time decreased significantly with increasing temperature.

    • Conclusions:

    • NBD-chloride is a more reactive and stable alternative to DNS-chloride for trypsin labeling.
    • NBD-chloride labeling of trypsin can be monitored spectrophotometrically.
    • Fluorescence depolarization provides insights into enzyme dynamics and temperature sensitivity.