Immuno-Northern Blotting: Detection of Modified RNA Using Gel Separation and Antibodies to Modified Nucleosides

Eikan Mishima; Takaaki Abe

doi:10.1007/978-1-4939-8808-2_13

Immuno-Northern Blotting: Detection of Modified RNA Using Gel Separation and Antibodies to Modified Nucleosides

Methods Mol Biol. 2019:1870:179-187. doi: 10.1007/978-1-4939-8808-2_13.

Authors

Eikan Mishima¹, Takaaki Abe²

Affiliations

¹ Division of Nephrology, Endocrinology, and Vascular Medicine, Tohoku University Graduate School of Medicine, Sendai, Japan.
² Department of Medical Science, Tohoku University Graduate School of Biomedical Engineering, Sendai, Japan. takaabe@med.tohoku.ac.jp.

PMID: 30539555
DOI: 10.1007/978-1-4939-8808-2_13

Abstract

Immuno-northern blotting is a method for detecting modified RNAs using gel separation and specific antibodies to modified nucleosides. This method was developed by combining two commonly used molecular biology techniques: western blotting and northern blotting. In this method, urea-polyacrylamide (or agarose) gel-separated RNAs are transferred to positively charged nylon membrane and then immune detection is performed with specific antibodies to modified nucleosides: such as 1-methyladenosine, N6-methyladenosine, and pseudouridine. This highly sensitive and relatively simple method, which uses widely available laboratory equipment, enables small laboratories to compare the abundance of modified nucleic acids across samples.

Keywords: 1-Methyladenosine; Modified nucleotide; N6-Methyladenosine; Pseudouridine; RNA modification; Small RNAs; tRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine / analogs & derivatives
Antibodies*
Blotting, Northern* / methods
Nucleosides*
RNA* / chemistry
RNA* / genetics

Substances

Antibodies
Nucleosides
1-methyladenosine
RNA
N-methyladenosine
Adenosine