Characterization of RNA-Binding Proteins Using Protein Microarrays

Guang Song; Johnathan Neiswinger; Heng Zhu

doi:10.1101/pdb.prot087973

Characterization of RNA-Binding Proteins Using Protein Microarrays

Cold Spring Harb Protoc. 2016 Oct 3;2016(10). doi: 10.1101/pdb.prot087973.

Authors

Guang Song¹, Johnathan Neiswinger¹, Heng Zhu²

Affiliations

¹ Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
² Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Center for High-Throughput Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287.

PMID: 27698236
DOI: 10.1101/pdb.prot087973

Abstract

RNA-binding proteins (RBPs), along with target RNA, play a vital role in the regulation of cellular processing and are especially important in gene transcription and posttranscriptional regulation. Here, we present a high-throughput method for rapid identification of RBPs for a given RNA using protein microarray technology. This protocol includes preparation of Cy5-labeled RNA probes, probe denaturing and refolding, and an RNA-binding assay as performed on a yeast protein microarray.

MeSH terms

Fungal Proteins / metabolism*
High-Throughput Screening Assays / methods
Microarray Analysis / methods*
Protein Array Analysis / methods*
Protein Binding
RNA / metabolism*
RNA-Binding Proteins / metabolism*

Substances

Fungal Proteins
RNA-Binding Proteins
RNA