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Uma Análise Pull-Down Quantitativa Otimizada de Proteínas Ligadoras de RNA Usando RNA Biotinilado Curto
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA

Uma Análise Pull-Down Quantitativa Otimizada de Proteínas Ligadoras de RNA Usando RNA Biotinilado Curto

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2,195 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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Aqui, apresentamos um método in vitro otimizado para descobrir, quantificar e validar interatores proteicos de sequências específicas de RNA, usando extrato proteico total de células humanas, esferas de estreptavidina revestidas com RNA biotinilado e análise por espectrometria de massas.

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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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