title: High-Throughput Quantitative Proteomics Enabled by Mass Defect-Based 12-Plex DiLeu Isobaric Tags.
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.
authors: School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI, 53705, USA.
authors: Dustin C
authors: DC
authors: Frost
authors: School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI, 53705, USA. lingjun.li@wisc.edu.|Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, WI, 53706, USA. lingjun.li@wisc.edu.
authors: Lingjun
authors: L
authors: Li
mesh_terms: D002853:Chromatography, Liquid; D007930:Leucine / Q000737:chemistry; D040901:Proteomics / Q000379:methods*
chemical_list: D007930:Leucine
keywords: DiLeu; High-resolution mass spectrometry; Isobaric labeling; Isobaric tag synthesis; Mass defect; Multiplexed quantitation; Quantitative proteomics