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Method | Open Access

Quantitation of nucleoprotein complexes by UV absorbance and Bradford assay

Jiang Chen1,2,3Hao Luo1,3Mei Tao1Zhongchuan Liu1Ganggang Wang1( )
Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences; Key Laboratory of Environmental Microbiology of Sichuan Province, Chengdu 610041, China
College of Life Sciences, Sichuan University, Chengdu 610064, China
University of Chinese Academy of Sciences, Beijing 100049, China
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Abstract

Despite the importance of studying nucleoprotein complexes, no appropriate method for quantifying them is available. Here, a UV absorbance method using the formula “Cmg/mL = 1.55A280 – 0.76A260” were applied to quantify nucleoprotein complexes. After modification using two paired A260 and A280 values, the UV-derived formula-based method could accurately quantify proteins in nucleoprotein complexes. Otherwise, by taking the target protein as a standard, the Bradford assay can accurately quantify proteins in nucleoprotein complexes without interference by nucleic acids. The above methods were successfully applied to measure the concentration of MtuP49-CTG complexes of Mycobacterium tuberculosis. In conclusion, both the Bradford assay and the UV-derived formula-based method were appropriate for quantifying proteins in nucleoprotein complexes, which may make contributions to explore the interactions between proteins and nucleic acids at the molecular level.

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Biophysics Reports
Pages 429-436
Cite this article:
Chen J, Luo H, Tao M, et al. Quantitation of nucleoprotein complexes by UV absorbance and Bradford assay. Biophysics Reports, 2021, 7(6): 429-436. https://doi.org/10.52601/bpr.2021.210028

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Received: 27 July 2021
Accepted: 26 November 2021
Published: 28 February 2022
© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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