biostudies-literatureUnknown97(41)Liu PKIsobaric labeling techniques are widely used in mass spectrometry-based quantitative proteomics to enable the simultaneous analysis of multiple samples. However, commercial isobaric tags are expensive due to complex synthesis and costly reagents, limiting their use in large-scale studies. Here, we introduce a novel, cost-effective diethylalanine-based isobaric reagent (DeAla), synthesized using diethylated alanine and β-alanine with <i>N</i>-hydroxysuccinimide. The DeAla tag offers several advantages, including improved peptide fragmentation, enhanced protein identification, and competitive pricing. We optimized labeling efficiency and collision energy parameters, demonstrating that DeAla-labeled peptides produce more backbone fragmentation ions and higher XCorr values compared to peptidesAnalytical chemistry22643-22650https://www.ebi.ac.uk/biostudies/studies/S-EPMC12547851biostudies-literature13-Plex DeAla Isobaric Reagents for High-Throughput Proteome Quantification.PMC12547851Nassar AGu TJWang DXu SWang ZLi LLiu PKChiang HYfalse13-Plex DeAla Isobaric Reagents for High-Throughput Proteome Quantification.Isobaric labeling techniques are widely used in mass spectrometry-based quantitative proteomics to enable the simultaneous analysis of multiple samples. However, commercial isobaric tags are expensive due to complex synthesis and costly reagents, limiting their use in large-scale studies. Here, we introduce a novel, cost-effective diethylalanine-based isobaric reagent (DeAla), synthesized using diethylated alanine and β-alanine with <i>N</i>-hydroxysuccinimide. The DeAla tag offers several advantages, including improved peptide fragmentation, enhanced protein identification, and competitive pricing. We optimized labeling efficiency and collision energy parameters, demonstrating that DeAla-labeled peptides produce more backbone fragmentation ions and higher XCorr values compared to peptides2025-01-01T00:00:00Z2025 Oct2026-06-04T16:22:14.11Z2026-06-02T03:08:00.462ZS-EPMC125478514105394010.1021/acs.analchem.5c03910