Label-Free Sample Preparation for Single-Cell Proteomics

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2024 Jun 21

PMID 38907142

Authors

David Hartlmayr

Claudia Ctortecka

Rupert Mayer

Karl Mechtler

Anjali Seth

Affiliations

Soon will be listed here.

Abstract

In recent years, single-cell proteomics (SCP) has become a valuable addition to other single-cell omics technologies for studying cellular heterogeneity. The amount of protein in a single cell is very limited, and in contrast to sequencing techniques, there are currently no means for protein amplification. Therefore, most single-cell proteomics approaches aim to maximize sample preparation efficiency while minimizing peptide loss. By reducing processing volumes to sub-microliters and avoiding manual transfer steps that could lead to peptide loss, peptide recovery, and the robustness of SCP workflows have been significantly improved. In this chapter, we describe a protocol for label-free SCP sample preparation using the cellenONE platform and the proteoCHIP LF 48 substrate prior to analysis with high-performance liquid chromatography-mass spectrometry.

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