Chapter title |
LC-MALDI-TOF/TOF for Shotgun Proteomics.
|
---|---|
Chapter number | 2 |
Book title |
Shotgun Proteomics
|
Published in |
Methods in molecular biology, April 2014
|
DOI | 10.1007/978-1-4939-0685-7_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0684-0, 978-1-4939-0685-7
|
Authors |
Fernández-Puente P, Mateos J, Blanco FJ, Ruiz-Romero C, Patricia Fernández-Puente, Jesús Mateos, Francisco J. Blanco M.D., Cristina Ruiz-Romero, Fernández-Puente, Patricia, Mateos, Jesús, Blanco, Francisco J., Ruiz-Romero, Cristina, Francisco J. Blanco |
Editors |
Daniel Martins-de-Souza |
Abstract |
Automated liquid chromatography tandem mass spectrometry (LC-MS/MS) is a well-established technique for identification of components from complex mixtures in shotgun proteomics experiments. Approaches involving the use of matrix-assisted laser desorption/ionization mass spectrometry (LC-MALDI-MS/MS) comprise the preparation of protein extracts, their enzymatic digestion, the separation of the resulting peptides by nanoscale liquid chromatography coupled to a collector that deposits the microfractions onto a MALDI plate, and finally the mass spectrometry analysis of the fractions. Using an LC-MALDI strategy, the rate of the collection of MS/MS data is decoupled from the chromatographic separation, thus allowing high-quality data which are often complementary to electrospray ionization (LC-ESI-MS/MS) techniques. |
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