Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Peptide Identification Using Tandem Mass Spectrometry

Peptide Identification Using Tandem Mass Spectrometry

Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...

Two-dimensional Gel Electrophoresis

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...

You might also read

Related Articles

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

Sort by

Same author

Influence of oven-drying pre-treatment on the metabolic profile of extracts from brewer's spent grains.

Frontiers in nutrition·2026

Same author

Evaluation of a high throughput multiparallel stirred bioreactor system using an apple cell line.

Scientific reports·2026

Same author

Differential modulation of tomato root exudates by <i>Streptomyces</i> strains underlies contrasting control of <i>Fusarium oxysporum</i> f. sp. <i>lycopersici</i>.

Frontiers in plant science·2026

Same author

Comparative metabolomic profiling of <i>Lupinus albus</i> and <i>L. angustifolius</i> harvest residues: exploring chemical diversity and valorization potential.

Frontiers in plant science·2025

Same author

Harnessing Apple Cell Suspension Cultures in Bioreactors for Triterpene Production: Transcriptomic Insights into Biomass and Triterpene Biosynthesis.

International journal of molecular sciences·2025

Same author

The effect of high-level dietary Laminaria digitata on the muscle proteome and metabolome of weaned piglets.

Research in veterinary science·2025

Search research articles

Related Experiment Video

Updated: Jun 23, 2026

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Published on: April 2, 2018

C-terminal sequence analysis of 2DE-separated proteins.

Bart Samyn¹, Kjell Sergeant, Jozef Van Beeumen

¹Laboratory of Protein Biochemistry and Protein Engineering, Department of Biochemistry, Physiology and Microbiology, Ghent University, K.L. Ledeganckstraat 35, B-9000, Gent, Belgium.

Methods in Molecular Biology (Clifton, N.J.)

|April 22, 2009

Summary

This study introduces a new method to identify protein C-terminal sequences using 2D gel electrophoresis and mass spectrometry. This approach systematically analyzes proteolytic processing, a common post-translational modification.

More Related Videos

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Related Experiment Videos

Last Updated: Jun 23, 2026

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome

Published on: April 2, 2018

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Area of Science:

Biochemistry
Proteomics

Background:

Post-translational modifications (PTMs) are crucial for protein function.
Protein truncations, particularly N- and C-terminal modifications, are common PTMs.
Systematic analysis of proteolytic processing events remains underexplored.

Purpose of the Study:

To develop and present a protocol for identifying the C-terminal sequences of proteins.
To enable systematic analysis of C-terminal proteolytic processing.
To investigate the effectiveness of the developed protocol using a model system.

Main Methods:

Two-dimensional polyacrylamide gel electrophoresis (2DE) for protein separation.
Cyanogen bromide cleavage to generate peptide mixtures.
Carboxypeptidase incubation to form characteristic ladders from original C-terminal fragments.
MALDI mass spectrometry for ladder readout.

Main Results:

A protocol was successfully developed for identifying protein C-terminal sequences after 2DE separation.
The method effectively identifies the original C-terminal fragment through ladder formation.
The protocol demonstrated effectiveness using proteins from Shewanella oneidensis as a model.

Conclusions:

The presented protocol offers a systematic approach to analyze C-terminal protein truncations.
This method enhances the study of proteolytic processing events.
The technique provides valuable insights into protein maturation and function.