Muscle Biopsy Sample Preparation and Proteomics Analysis Based on UHPLC-MS/MS.

Abstract

Proteomics analysis is crucial for understanding the molecular mechanisms underlying muscle adaptations to different types of exercise, such as concentric and eccentric training. Traditional methods like two-dimensional gel electrophoresis and standard mass spectrometry have been used to analyze muscle protein content and modifications. This protocol details the preparation of muscle samples for proteomics analysis using ultra-high-performance liquid chromatography (UHPLC). It includes steps for muscle biopsy collection, protein extraction, digestion, and UHPLC-based analysis. The UHPLC method offers high-resolution separation of complex protein mixtures, providing more detailed and accurate proteomic profiles compared to conventional techniques. This protocol significantly enhances sensitivity, reproducibility, and efficiency, making it ideal for comprehensive muscle proteomics studies. Key features • Developed for analyzing muscle adaptations in response to concentric and eccentric training, applicable to various physiology exercise studies. • Utilizes UHPLC-MS/MS for high-resolution separation and detailed proteomic profiling. • Requires access to advanced UHPLC-MS/MS equipment and muscle biopsy collection tools. • The protocol can be completed within one week, including sample preparation and analysis.