Fig. 1: sCIP-TMT allows for more efficient sample preparation with less sample-to-sample variance.
From: Functionalizing tandem mass tags for streamlining click-based quantitative chemoproteomics
A Workflow currently used for profiling cysteines in which samples are labeled with either iodoacetamide-desthiobiotin (IA-DTB) or iodoacetamide alkyne (IAA) and conjugated to biotin azide via copper-catalyzed azide-alkyne cycloaddition (CuAAC or ‘click’). After sample cleanup, using single-pot solid-phase enhanced sample-preparation (SP3), as illustrated here, or other decontamination methodologies, the samples are then subjected to sequence-specific proteolytic digest, isobaric labeling, avidin enrichment sample pooling, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. B Our envisioned sCIP-TMT workflow in which fully functionalized isobaric, biotin- and azide-containing reagents allow for early-stage sample pooling directly after click conjugation. Subsequently, the labeled samples can be processed and analyzed following the established sample preparation workflow.
