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

Internal sequences from proteins digested in polyacrylamide gels

P Jenö1, T Mini, S Moes

  • 1Department of Biochemistry, Biozentrum of the University of Basel, Switzerland.

Analytical Biochemistry
|January 1, 1995
PubMed
Summary
This summary is machine-generated.

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

Long-Range Transverse-Momentum Correlations and Radial Flow in Pb-Pb Collisions at the LHC.

Physical review letters·2026
Same author

Search for Quasiparticle Scattering in the Quark-Gluon Plasma with Jet Splittings in pp and Pb-Pb Collisions at sqrt[s_{NN}]=5.02  TeV.

Physical review letters·2025
Same author

First Measurement of A=4 Hypernuclei and Antihypernuclei at the LHC.

Physical review letters·2025
Same author

Long-term follow-up after ureteral reimplantation in children: a 12-year analysis.

Pediatric surgery international·2025
Same author

Probing Strangeness Hadronization with Event-by-Event Production of Multistrange Hadrons.

Physical review letters·2025
Same author

Measurements of Chemical Potentials in Pb-Pb Collisions at sqrt[s_{NN}]=5.02  TeV.

Physical review letters·2024

A new method simplifies protein digestion for mass spectrometry analysis. This technique avoids blotting and yields extensive peptide fragments from Coomassie blue-stained proteins in gels.

Area of Science:

  • Biochemistry
  • Proteomics
  • Analytical Chemistry

Background:

  • Proteolytic digestion is crucial for protein identification via mass spectrometry.
  • In-gel digestion of stained proteins can be complicated by disulfide bond formation.
  • Existing methods often require additional steps like blotting.

Purpose of the Study:

  • To develop a simplified method for proteolytic digestion of Coomassie blue-stained proteins directly within a polyacrylamide gel matrix.
  • To enable efficient generation of internal peptide sequences for protein identification.

Main Methods:

  • Proteins stained with Coomassie blue were reduced and alkylated using dithiothreitol and iodoacetamide in the presence of sodium dodecyl sulfate.
  • The modified proteins were then digested using the endoproteinase LysC.

Related Experiment Videos

  • This process was performed directly on proteins within the polyacrylamide gel.

    • Main Results:

    • The method effectively suppresses disulfide bond formation during in-gel digestion.
    • Extensive proteolysis was achieved, comparable to solution digests.
    • Internal sequence data was successfully generated from low microgram quantities of excised proteins.
    • No blotting step was required for internal sequence generation.

    Conclusions:

    • This simple, effective method allows for direct proteolytic digestion of Coomassie blue-stained proteins in polyacrylamide gels.
    • The technique streamlines the process for generating internal sequence data, facilitating protein identification.
    • It offers an advantageous alternative to existing methods by eliminating the need for blotting.