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TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis.

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.

SUBMITTER: Suzuki-Hatano S

PROVIDER: S-EPMC7442215 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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TMT Sample Preparation for Proteomics Facility Submission and Subsequent Data Analysis.

Suzuki-Hatano Silveli S Tsai Ang-Chen AC Daugherty Audrey A Pacak Christina A CA

Journal of visualized experiments : JoVE 20200608 160

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 consiste ...[more]

PMID: 32568242

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