Faculty, Staff and Student Publications

Publication Date

7-1-2025

Journal

STAR Protocols

DOI

10.1016/j.xpro.2025.103924

PMID

40601427

PMCID

PMC12268536

PubMedCentral® Posted Date

7-1-2025

PubMedCentral® Full Text Version

Post-print

Abstract

tRNA modifications are highly dynamic and play crucial roles in regulating gene expression. Here, we present an assay for analyzing N2, N2-dimethylguanosine (m22G) modifications on tRNAs using an antibody-based detection approach. We describe steps for isolating tRNAs from human cells via ion-exchange chromatography, capturing specific tRNA iso-decoders on streptavidin magnetic beads, and detecting modifications through immuno-northern blotting. This approach enables researchers to investigate other tRNA modifications using specific antibodies or downstream approaches such as mass spectrometry.

Keywords

Cell Biology, Molecular Biology, Molecular/Chemical Probes

Published Open-Access

yes

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