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

Author

Marín-Vicente C, Calvo E, Rodríguez JM, Villa Del Campo C, Sierra R, Végvári Á, Zubarev RA, Torres M, and Vázquez J

Subjects
Humans, Mass Spectrometry methods, Flow Cytometry methods, Proteome analysis, Animals, Isotope Labeling methods, Myocytes, Cardiac metabolism, Myocytes, Cardiac cytology, Staining and Labeling methods, Proteomics methods, Single-Cell Analysis methods
Abstract

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., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Published
2024
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