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Related Concept Videos

MALDI-TOF Mass Spectrometry01:19

MALDI-TOF Mass Spectrometry

Mass spectrometry is a powerful characterization technique that can identify and separate a wide variety of compounds ranging from chemical to biological entities, based on their mass-to-charge ratio (m/z). The instruments that allow this detection, known as mass spectrometers, have three components: an ion source, a mass analyzer, and a detector. These spectrometers differ based on the nature of their ion source and analyzers.Matrix-assisted laser desorption ionization (MALDI) is a commonly...
Peptide Identification Using Tandem Mass Spectrometry01:33

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 Electrophoresis01:22

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...
Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
Matrix-Assisted Laser Desorption Ionization (MALDI)01:08

Matrix-Assisted Laser Desorption Ionization (MALDI)

Matrix-assisted laser desorption ionization (MALDI) is a powerful analytical technique used in mass spectrometry. It enables the identification and characterization of various biomolecules, including proteins, peptides, nucleic acids, and carbohydrates. MALDI is an ionization technique, widely employed in biological and medical research, as well as in fields like pharmacology and biochemistry.The analyte of interest, a biomolecule or a mixture of biomolecules, is mixed with a suitable matrix...
Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...

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Related Experiment Video

Updated: Jun 25, 2026

Two-dimensional Gel Electrophoresis Coupled with Mass Spectrometry Methods for an Analysis of Human Pituitary Adenoma Tissue Proteome
12:34

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

Published on: April 2, 2018

Classical proteomics: two-dimensional electrophoresis/MALDI mass spectrometry.

Ursula Zimny-Arndt1, Monika Schmid, Renate Ackermann

  • 1Max Planck Institute for Infection Biology, Berlin, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|February 26, 2009
PubMed
Summary

Proteomics advancements enable detailed protein analysis using two-dimensional electrophoresis (2-DE) and mass spectrometry (MS). This combination provides high-resolution separation and sensitive identification of protein species for structural elucidation.

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Last Updated: Jun 25, 2026

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

Published on: April 2, 2018

Proteomic Profiling of Macrophages by 2D Electrophoresis
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Proteomic Profiling of Macrophages by 2D Electrophoresis

Published on: November 4, 2014

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

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

Published on: April 1, 2017

Area of Science:

  • Proteomics
  • Biochemistry
  • Molecular Biology

Background:

  • The field of proteomics has experienced rapid technological advancements in the past decade.
  • These developments aim to comprehensively analyze proteins within biological systems.
  • Two-dimensional electrophoresis (2-DE) offers high-resolution separation of complex protein mixtures.

Purpose of the Study:

  • To present a robust protocol for detailed protein analysis.
  • To combine high-resolution separation with sensitive identification methods.
  • To elucidate protein species structures, including post-translational modifications and genetic variations.

Main Methods:

  • Two-dimensional electrophoresis (2-DE) for high-resolution protein separation (>10,000 spots).
  • Sensitive protein identification using peptide mass fingerprinting and MS/MS sequencing.
  • Off-line measurement by MALDI-TOFTOF-MS for repeated sample analysis and comprehensive structural data.

Main Results:

  • 2-DE successfully separates proteins based on charge and mass, resolving various protein species.
  • Peptide mass fingerprinting combined with MS/MS provides high sequence coverage for protein identification.
  • MALDI-TOFTOF-MS enables repeated measurements, yielding more complete protein structure information.

Conclusions:

  • The presented protocol, integrating 2-DE and MALDI-TOFTOF-MS, is effective for detailed proteomic analysis.
  • This approach allows for the structural elucidation of individual protein species.
  • The methodology is foundational for advancing our understanding of complex biological systems at the protein level.