Protocols for Proteomic Analysis: Isolation, Solubilization and Hydrolysis by Proteases

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Published: Jun 20, 2025
DOI: https://doi.org/10.18097/BMCRM00277
Keywords:
proteomics; protein solubilization; highly specific protease; protocol

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E.N. Obukhova
Skolkovo Institute of Science and Technology, 30c1 Bolshoi Boulevard, Moscow, 121205 Russia
E.V. Khryapova
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119435 Russia
S.E. Novikova
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119435 Russia
L.Sh. Kazieva
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119435 Russia
O.V. Tikhonova
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119435 Russia
V.G. Zgoda
Institute of Biomedical Chemistry, 10 Pogodinskaya str., Moscow, 119435 Russia

Abstract

High-throughput studies of protein composition of biological samples have become routine and are used practically in all areas of life sciences. Modern proteomics methods allow reliable identification and quantification of thousands of proteins in a single experiment. The standard procedure for proteomic analysis includes the following steps: 1. isolation and solubilization of proteins, their hydrolysis by proteases; 2. analysis of the resulting peptides by high-performance liquid chromatography with mass spectrometric detection; 3. bioinformatics and statistical processing of the results. This paper presents protocols of the first stage of proteomic analysis, i.e. sample preparation, which are routinely used in the Laboratory of Systems Biology of the Institute of Biomedical Chemistry.

Article Details

How to Cite
Obukhova, E., Khryapova, E., Novikova, S., Kazieva, L., Tikhonova, O., & Zgoda, V. (2025). Protocols for Proteomic Analysis: Isolation, Solubilization and Hydrolysis by Proteases. Biomedical Chemistry: Research and Methods, 8(2), e00277. https://doi.org/10.18097/BMCRM00277
Issue
Vol. 8 No. 2 (2025)
Section
PROTOCOLS OF EXPERIMENTS, USEFUL MODELS, PROGRAMS AND SERVICES

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