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An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA
JoVE 杂志
生物化学
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JoVE 杂志 生物化学
An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA

An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA

2,201 Views

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07:55 min

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February 17, 2023

DOI:

10.3791/64923-v

DOI:
10.3791/64923-v

•

07:55 min
February 17, 2023

•

40 Views
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1Center for Human Technology,Italian Institute of Technology (IIT), 2Department of Experimental Oncology,European Institute of Oncology (IEO), 3Department of Biology and Biotechnologies ‘Charles Darwin’,Sapienza University of Rome

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Here, we present an optimized in vitro method to uncover, quantify, and validate protein interactors of specific RNA sequences, using total protein extract from human cells, streptavidin beads coated with biotinylated RNA, and mass spectrometry analysis.

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Crua Asensio, N., Rizzieri, S., Cuomo, A., Tartaglia, G. G., Zacco, E. An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA. J. Vis. Exp. (192), e64923, doi:10.3791/64923 (2023).

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