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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...

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Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

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Published on: July 1, 2014

Reversed-phase isolation of peptides.

W J Henzel¹, J T Stults

¹Genentech, Inc., South San Francisco, California.

Current Protocols in Molecular Biology

|February 12, 2008

Summary

This summary is machine-generated.

This study details protocols for high-performance liquid chromatography (HPLC) to separate peptides. It covers methods for various sample quantities, enabling downstream analysis like mass spectrometry.

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Sample Preparation for Endopeptidomic Analysis in Human Cerebrospinal Fluid

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Sample Preparation for Endopeptidomic Analysis in Human Cerebrospinal Fluid

Sample Preparation for Endopeptidomic Analysis in Human Cerebrospinal Fluid

Published on: December 4, 2017

Area of Science:

Biochemistry
Analytical Chemistry
Chromatography

Background:

Reversed-phase high-performance liquid chromatography (RP-HPLC) is a key technique for peptide separation.
Effective separation of peptides is crucial for subsequent analyses, such as automated sequencing and mass spectrometry.

Purpose of the Study:

To provide detailed protocols for peptide separation using reversed-phase HPLC.
To describe methods applicable to a wide range of peptide quantities, from <5 pmol to 5–500 pmol.
To facilitate the analysis of complex peptide mixtures.

Main Methods:

Utilizing reversed-phase HPLC with hydrophobic stationary phases.
Employing gradient elution with increasing organic solvent concentrations.
Describing protocols for narrow-bore (2-mm-i.d.) and microbore (1-mm-i.d.) capillary HPLC columns.
Specifying gradient flow rates for capillary HPLC columns (3–5 µL/min).

Main Results:

Established protocols for separating peptides in quantities ranging from <5 pmol to 5–500 pmol.
Demonstrated the feasibility of peptide separation using narrow-bore and microbore HPLC columns.
Enabled the analysis of components present in small samples prior to automated sequencing.

Conclusions:

The provided HPLC protocols enable efficient peptide separation across diverse sample amounts.
These methods are essential for preparing samples for advanced analytical techniques like matrix-assisted laser desorption/ionization mass spectrometry and capillary electrophoresis.
The study enhances the capability for sensitive peptide analysis in biological and chemical research.