Electrophoretic mobility shift assays (EMSAs) for in vitro detection of protein-nucleic acid interactions.

Gene Expression Molecular Biology Molecular/Chemical Probes Protein Biochemistry

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
13 Jun 2024
Historique:
received: 19 01 2024
revised: 11 04 2024
accepted: 23 05 2024
medline: 14 6 2024
pubmed: 14 6 2024
entrez: 14 6 2024
Statut: aheadofprint

Résumé

Protein-nucleic acid interactions drive some of the most important physiological events in cells. Here, we present a protocol for detecting protein-DNA or protein-RNA interactions in vitro. We describe steps for labeling nucleic acid species and electrophoretic mobility shift assays (EMSAs). This protocol can be used to confirm suspected in vivo interactions using recombinantly expressed/purified proteins of interest and a nucleic acid substrate. It can further be used to investigate mutations that can disrupt interaction or compensatory mutations that restore it. For complete details on the use and execution of this protocol, please refer to Mansouri-Noori et al.

Identifiants

pubmed: 38875114
pii: S2666-1667(24)00293-4
doi: 10.1016/j.xpro.2024.103128
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

103128

Informations de copyright

Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Auteurs

Farnaz Mansouri-Noori (F)

Department of Biology, York University, Toronto, ON M3J 1P3, Canada. Electronic address: farnaz.mansouri@gmail.com.

Mark A Bayfield (MA)

Department of Biology, York University, Toronto, ON M3J 1P3, Canada. Electronic address: bayfield@yorku.ca.

Classifications MeSH