•Live cell imaging-based assays can be used to study assembly and intra-nuclear location of protein complexes that regulate RNA polymerase II.•This article describes use of AP-FRET to study assembly of the Elongin ubiquitin ligase.•It also describes laser microirradiation-based assays used to study its recruitment to regions of localized DNA damage. Elongin A binds to Elongins B and C to form the RNA polymerase II transcription elongation factor Elongin. It also functions as the substrate recognition subunit of a ubiquitin ligase that is formed by binding of Elongin to Cullin protein CUL5 and RING finger protein RBX2 and that targets RNA polymerase II for ubiquitination. In this article, we describe use of acceptor photobleaching fluorescence resonance energy transfer (AP-FRET) and laser microirradiation-based assays to study regulated assembly of the Elongin ubiquitin ligase and its recruitment to regions of localized DNA damage.