Rapid Isolation of Lysosomes (LysoIP) from iPSC-Derived Neurons for Proteomics, with Options for Lipidomics/Metabolomics v1

This protocol describes rapid immunopurification of lysosomes (LysoIP) from cultured iNeurons expressing a lysosomal HA tag (e.g., TMEM192-3xHA). The workflow is optimized forproteome analysis from one 10-cm dish (~8–10×10^6 iNeurons)and can be scaled by pooling multiple dishes for lipidomics/metabolomics. The method relies on cold, rapid handling; gentle homogenization; anti-HA magnetic capture; stringent washes; and extraction compatible with downstream mass spectrometry.