Protein sample preparation for tissue distribution study.

Autor: Cheng L; Center for Life Sciences, School of Life Sciences, Yunnan University, Kunming, China.; Institute of Biophysics, Chinese Academy of Sciences, Beijing, China., Xu Y; School of Basic Medical Sciences, Southwest Medical University, Luzhou, China.; Institute of Biophysics, Chinese Academy of Sciences, Beijing, China., Zhu K; Huazhong University of Science and Technology, Wuhan, China., Liang B; Center for Life Sciences, School of Life Sciences, Yunnan University, Kunming, China., Zhang S; Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.; Institute of Infectious Diseases, Beijing Key Laboratory of Emerging Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, China.; Beijing Institute of Infectious Diseases, Beijing, China.; National Center for Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, China., Liu P; School of Basic Medical Sciences, Southwest Medical University, Luzhou, China.; Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.; University of Chinese Academy of Sciences, Beijing, China.
Jazyk: angličtina
Zdroj: Proteomics. Clinical applications [Proteomics Clin Appl] 2023 Mar; Vol. 17 (2), pp. e2200088. Date of Electronic Publication: 2022 Nov 16.
DOI: 10.1002/prca.202200088
Abstrakt: Purpose: The distribution and expression level of a protein among animal tissues is indicative of its possible roles. It is important to establish a generally applicable method to prepare protein samples with high-quality and achieve near 100% recovery of proteins from animal tissues.
Experimental Design: During preparation, to sufficiently dissolve and maintain stability of almost all proteins from tissues, as well as to avoid most contaminations affecting protein detection, 2×SDS Sample Buffer, sonication and trichloroacetic acid precipitation are applied.
Results: Here we provide a relatively simple, reproducible, and broadly applicable method for studying protein distribution in most tissues, in which the issues resulting from protein degradation and modification during sample preparation and assay interference by other cellular components like neutral lipids and glycogen could be overcome. Furthermore, this method represents the protein content by equal wet tissue mass, which is a better means to present the expression level of a protein in various tissues. High-quality protein samples from almost all tissues could be prepared.
Conclusions and Clinical Relevance: The samples produced are amenable to tissue distribution analysis by Western blotting and for silver/Coomassie staining, proteomics, and other protein analyses, which would contribute to potential biomarkers or treatments for a disease.
(© 2022 Wiley-VCH GmbH.)
Databáze: MEDLINE
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