Ultraviolet Photochemical Cross-Linking to Detect RNA-Binding Proteins
Adapted from RNA: A Laboratory Manual, by Donald C. Rio, Manuel Ares Jr, Gregory J. Hannon, and Timothy W. Nilsen. CSHL Press, Cold Spring Harbor, NY, USA, 2011.Abstract
RNA is photoreactive on exposure to ultraviolet (UV) light in the 250- to 270-nm range. On UV treatment, RNA bases absorb energy to generate free radicals that can covalently attach to nearby amino acid residues in RNA-bound proteins. UV cross-linking experiments have been extensively used to identify and characterize RNA-binding proteins. The method described here involves the use of 32P-labeled RNA and crude extracts or purified proteins.
- © 2013 Cold Spring Harbor Laboratory Press
