Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides.

ABSTRACT: The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB) analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A), N6-methyladenosine (m6A), pseudouridine, and 5-methylcytidine (m5C) showed different modifications in a variety of RNAs from various specie

SUBMITTER: Mishima E 

PROVIDER: S-EPMC4659547 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature