Many biological systems are composed of diverse single cells. This diversity necessitates functional and molecular single-cell analysis. Single-cell protein analysis has long relied on affinity reagents, but emerging mass-spectrometry methods (either label-free or multiplexed) have enabled quantifying over 1,000 proteins per cell while simultaneously increasing the specificity of protein quantification. Isobaric carrier based multiplexed single-cell proteomics is a scalable, reliable, and cost-effective method that can be fully automated and implemented on widely available equipment. It uses inexpensive reagents and is applicable to any sample that can be processed to a single-cell suspension. Here we describe an automated SCoPE2 workflow that allows analyzing about 200 single cells per 24 hours using only standard commercial equipment.
The presentations below describe sample preparation and data analysis for the latest version of Single Cell ProtEomics by Mass Spectrometry (SCoPE-MS), SCoPE2
- Design of single-cell proteomics experiments using SCoPE2
- Sample preparation for single-cell MS analysis by SCoPE2
- High-throughput single-cell proteomics quantifies the emergence of macrophage heterogeneity