Sample Preparation for Proteomic Analysis of Isolated Mitochondria and Whole-Cell Extracts
Output Details
Description
Mass spectrometry-based proteomics relies on precise sample preparation to ensure reliable analysis of proteomic changes, including post-translational modifications (PTMs). This protocol outlines a streamlined workflow for processing isolated mitochondria (or other organelles) and whole-cell extracts from cultured cells or mouse tissues. Key steps include robust lysis with 2% SDS, high-energy sonication, and protein capture on S-trap columns to remove interfering substances. Optimized trypsin/Lys-C digestion and sequential peptide elution enable high-quality mass spectrometry data acquisition. This method is adaptable for diverse proteomic and PTM studies.
Identifier (DOI)
10.17504/protocols.io.3byl4wrj8vo5/v1
