Chapter title |
Peptide Immunoaffinity Enrichment with Targeted Mass Spectrometry: Application to Quantification of ATM Kinase Phospho-Signaling
|
---|---|
Chapter number | 15 |
Book title |
ATM Kinase
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6955-5_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6953-1, 978-1-4939-6955-5
|
Authors |
Jeffrey R. Whiteaker, Lei Zhao, Regine M. Schoenherr, Jacob J. Kennedy, Richard G. Ivey, Amanda G. Paulovich |
Editors |
Sergei V. Kozlov |
Abstract |
Peptide immunoaffinity enrichment coupled with targeted mass spectrometry is a quantitative approach for the robust and reproducible quantification of peptide analytes. The approach is capable of multiplexed quantification of peptides, including posttranslational modifications such as phosphorylation. Anti-peptide antibodies are used to enrich analytes and heavy stable isotope-labeled standards. The enriched peptides are directly measured by multiple reaction monitoring (MRM), a well-characterized quantitative mass spectrometry-based method. Quantification is performed by measuring the analyte (light) peptide response relative to the heavy standard, which is spiked at a known concentration. Here, we describe the methodology for multiplexed measurement of phosphorylated peptides on the ATM kinase and their nonmodified peptide analogs in cellular lysates. The method provides quantitative measurements of phospho-signaling and can be extended to a number of other phosphopeptides and sample types. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 15 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Master | 3 | 20% |
Student > Ph. D. Student | 2 | 13% |
Student > Bachelor | 2 | 13% |
Researcher | 2 | 13% |
Unspecified | 1 | 7% |
Other | 0 | 0% |
Unknown | 5 | 33% |
Readers by discipline | Count | As % |
---|---|---|
Chemistry | 3 | 20% |
Biochemistry, Genetics and Molecular Biology | 2 | 13% |
Medicine and Dentistry | 2 | 13% |
Agricultural and Biological Sciences | 2 | 13% |
Unspecified | 1 | 7% |
Other | 0 | 0% |
Unknown | 5 | 33% |