| Chapter title |
High-Throughput Quantitative Proteomics Enabled by Mass Defect-Based 12-Plex DiLeu Isobaric Tags
|
|---|---|
| Chapter number | 10 |
| Book title |
Quantitative Proteomics by Mass Spectrometry
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3524-6_10 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3522-2, 978-1-4939-3524-6
|
| Authors |
Frost, Dustin C, Li, Lingjun, Dustin C. Frost, Lingjun Li |
| Editors |
Salvatore Sechi |
| Abstract |
Isobaric labeling has become a popular technique for high-throughput, mass spectrometry (MS)-based relative quantification of peptides and proteins. However, widespread use of the approach for large-scale proteomics applications has been limited by the high cost of commercial isobaric tags. To address this, we have developed our own N,N-dimethyl leucine (DiLeu) multiplex isobaric tags as a cost-effective alternative that can be synthesized with ease using readily available isotopic reagents. When paired with high-resolution tandem mass (MS (n) ) acquisition, mass defect-based DiLeu isobaric tags allow relative quantification of up to twelve samples in a single liquid chromatography (LC)-MS(2) experiment. Herein, we present detailed methods for synthesis of 12-plex DiLeu isobaric tags, labeling of complex protein digest samples, analysis by high-resolution nanoLC-MS (n) , and processing of acquired data. |
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