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

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.