Italian Institute of Technology (IIT) 1 article published in JoVE Biochemistry An Optimized Quantitative Pull-Down Analysis of RNA-Binding Proteins Using Short Biotinylated RNA Núria Crua Asensio1, Stefano Rizzieri2, Alessandro Cuomo2, Gian Gaetano Tartaglia1,3, Elsa Zacco1 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 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.