Chapter title |
Biomarker verification using selected reaction monitoring and shotgun proteomics.
|
---|---|
Chapter number | 20 |
Book title |
Shotgun Proteomics
|
Published in |
Methods in molecular biology, April 2014
|
DOI | 10.1007/978-1-4939-0685-7_20 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0684-0, 978-1-4939-0685-7
|
Authors |
Castro-Gamero AM, Izumi C, Rosa JC, Angel Mauricio Castro-Gamero, Clarice Izumi, José César Rosa, Castro-Gamero, Angel Mauricio, Izumi, Clarice, Rosa, José César |
Editors |
Daniel Martins-de-Souza |
Abstract |
Shotgun proteomics (liquid chromatography-electrospray ionization-mass spectrometry, LC-ESI-MS/MS) has dominated the strategies for global protein expression in subcells, cells, tissues, and whole organisms with several types of approaches, as isobaric tags for relative and absolute quantification (iTRAQ), isotope-coded affinity tags (ICAT), or stable isotope labeling using amino acids in cell culture (SILAC) and non-labeling (label free) methods. Shotgun proteomics practically replaced the classical 2D gel electrophoresis. Selected reaction monitoring (SRM), also denominated multiple reaction monitoring (MRM), is a targeted quantitative technology that uses a complex mixture of tryptic peptides that can be selectively detected by liquid chromatography coupled to electrospray triple-quadrupole mass spectrometer; this system can select precursor ions in combination with their correspondent product ions during collision-induced dissociation to produce specific detection related to a particular protein. Here we describe protocols that are efficient to produce a complete enzymatic trypsin digestion from complex biological matrices and concomitant material to be used for LC-SRM-MS and LC-ESI-MS/MS (labeled or label free). |
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Mendeley readers
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Student > Ph. D. Student | 8 | 25% |
Researcher | 6 | 19% |
Student > Master | 5 | 16% |
Student > Postgraduate | 3 | 9% |
Other | 1 | 3% |
Other | 2 | 6% |
Unknown | 7 | 22% |
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Agricultural and Biological Sciences | 6 | 19% |
Chemical Engineering | 2 | 6% |
Neuroscience | 2 | 6% |
Medicine and Dentistry | 2 | 6% |
Other | 6 | 19% |
Unknown | 8 | 25% |