Work

TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis

Year: 2020

Type: article

Abstract: Proteomic technologies are powerful methodologies that can aid our understanding of mechanisms of action in biological systems by providing a global view of the impact of a disease, treatment, or other condition on the proteome as a whole. This report provides a detailed protocol for the extraction, quantification, precipitation, digestion, labeling, and subsequent data analysis of protein samples. Our optimized TMT labeling protocol requires a lower tag-label concentration and achieves consistently reliable data. We have used this protocol to evaluate protein expression profiles in a variety of mouse tissues (i.e., heart, skeletal muscle, and brain) as well as cells cultured in vitro. In addition, we demonstrate how to evaluate thousands of proteins from the resulting dataset.

Source: Journal of Visualized Experiments

DOI: 10.3791/60970

Authors: Silveli Suzuki-Hatano, Ang-Chen Tsai, Audrey Daugherty, Christina A. Pacak

Institution: University of Florida

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: bronze

Funders: National Institutes of Health, International Centre for Bamboo and Rattan, National Heart, Lung, and Blood Institute

Award: Mechanisms and Treatment of Cardiac and Skeletal Muscular Dysfunction in Barth Syndrome