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A Sample Preparation Procedure for Isobaric Labeling-Based Single-Cell Proteomics

  • Consuelo Marín-Vicente,
  • Enrique Calvo,
  • José Manuel Rodríguez,
  • Cristina Villa del Campo,
  • Rocío Sierra,
  • Ákos Végvári,
  • Roman A. Zubarev,
  • Miguel Torres,
  • Jesús Vázquez

摘要

Mass spectrometry-based proteomics has traditionally been limited by the amount of input material for analysis. Single-cell proteomics has emerged as a challenging discipline due to the ultra-high sensitivity required. Isobaric labeling-based multiplex strategies with a carrier proteome offer an approach to overcome the sensitivity limitations. Following this as the basic strategy, we show here the general workflow for preparing cells for single-cell mass spectrometry-based proteomics. This protocol can also be applied to manually isolated cells when large cells, such as cardiomyocytes, are difficult to isolate properly with conventional fluorescence-activated cell sorting (FACS) sorter methods.