SP3-C18 sample processing for mass spectrometry based proteomics

This protocol is adapted from the original protocol published in the “Single-pot, solid-phase-enhanced sample preparation for proteomics experiments” by Hughes et al. (2019) which describes the SP3 (single-pot, solid-phase-enhanced) protocol, which uses paramagnetic beads and a hydrophilic interaction mechanism to bind, wash and elute proteins in a one-tube workflow, enabling efficient, near-lossless processing of diverse protein samples (including very small amounts) for downstream mass spectrometry-based proteomics. 

This protocol uses SP3 beads for protein extraction and detergent removal, combined with the C18 protocol for peptide desalting, adapted from  "Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips" by et al. (2007).