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

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2,207 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
, , , ,
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

Summary

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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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