Related Experiment Video
Updated: Jul 5, 2026
09:35
Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
Published on: April 1, 2017
Analysis of protein composition using multidimensional chromatography and mass spectrometry
Andrew J Link1, Jennifer L Jennings1, Michael P Washburn2
1Vanderbilt University School of Medicine, Nashville, Tennessee.
Current Protocols in Protein Science
|April 23, 2008
Summary
Automated liquid chromatography and mass spectrometry identify numerous proteins in complex mixtures. This advanced technique bypasses limitations of gel electrophoresis for direct solution-based protein identification.
Area of Science:
- Proteomics
- Analytical Chemistry
- Biochemistry
Background:
- Gel electrophoresis and in-gel digestion are common but limited methods for protein identification.
- Analyzing complex protein mixtures directly in solution presents significant challenges.
Purpose of the Study:
- To present an advanced method for identifying a large number of proteins from complex biological samples.
- To overcome the limitations associated with traditional gel-based proteomics techniques.
Main Methods:
- Multidimensional liquid chromatography was employed for peptide separation.
- Tandem mass spectrometry was utilized for peptide analysis.
- Automated database searching was coupled with mass spectrometry data for protein identification.
Main Results:
- The combined methods successfully identified a large number of proteins from complex samples.
- The approach directly analyzed protein mixtures in solution, avoiding gel electrophoresis steps.
Conclusions:
- Multidimensional liquid chromatography and tandem mass spectrometry offer a powerful alternative for comprehensive protein identification.
- This solution-based approach enhances the efficiency and scope of proteomic analysis in complex biological systems.
Related Concept Videos
01:33Peptide 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...
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...
01:21Tandem 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...
01:21Mass 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...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
01:19MALDI-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...
01:33Proteomics
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term proteomics...