期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 589, 期 -, 页码 197-203出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2021.12.026
关键词
Shotgun proteomics; Sodium dodecyl sulfate; S-Trap digestion; Pellet S-Trap digestion
资金
- Korea Basic Science Institute, Korea [C170100]
- National Cancer Center, Korea [NCC-1810861]
- National Research Council of Science & Technology (NST), Republic of Korea [C170100] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Protein extraction and digestion are crucial steps in proteomics research. The use of SDS buffer enables effective analysis of diverse proteins. This study evaluated the use of the S-Trap digestion method combined with SDS buffer extraction and compared it to the traditional In solution digestion method. The results showed that S-Trap digestion with SDS buffer increased the number of identified proteins, particularly those located inside multilayer membranes.
Protein extraction and digestion are important analytical steps in the study of proteomics. The use of sodium dodecyl sulfate (SDS) buffer makes it possible to effectively analyze various proteins. Its use was evaluated using the S-Trap digestion method and compared to the traditional In solution digestion method. Differences in protein composition were examined for each protein preparation method. S-Trap digestion followed by SDS buffer extraction clearly increased the number of identified proteins, including more mitochondrial and membrane-related proteins. The S-Trap digestion method with 5% SDS buffer was applied to the pellet remaining from the removal of RIPA buffer-soluble proteins, which identified more extracellular space proteins than the conventional S-Trap digestion method. S-Trap digestion of the pellet was particularly advantageous for identifying proteins located inside multilayer membranes. (c) 2021 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
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