Protocol 1: Lysosomal Immunoprecipitation (LysoIP) From Cells and Tissues for Metabolomic and Proteomic Analyses​

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To study the composition of lysosomes, the dynamics of their contents, and their roles in health and disease, we use a lysosomal immunoprecipitation method known as LysoIP. This technique enables the isolation of intact lysosomes from both cultured cells and mouse tissues. The method relies on the expression of a lysosome-localized 3xHA epitope tag (LysoTag) and a simple, yet robust organelle immunoprecipitation workflow. Isolated lysosomes are extracted with optimized buffers, allowing for the efficient retrieval of lysosomal proteins, polar metabolites, and lipids. The process is fully compatible with downstream liquid chromatography and mass spectrometry (LC-MS) analyses.

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The detailed step-by-step protocol is available in the attached PDF.
👉 [Click here to view or download the full protocol. (PDF)]

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