Isolation and Immunodetection of Enzymatic DNA–Protein Crosslinks by RADAR Assay

Megan Perry, Gargi Ghosal

Research output: Chapter in Book/Report/Conference proceedingChapter


DNA–protein crosslinks (DPCs) are steric hindrances to DNA metabolic processes and the removal and repair of DPCs is a rapidly evolving area of research. A critical component of deciphering this repair pathway is developing techniques that detect and quantify specific types of DPCs in cells. Here we describe a protocol for direct detection of enzymatic DPCs from mammalian cells—the RADAR assay. The method involves isolating genomic DNA and DPCs from cells and binding them to nitrocellulose membrane with a vacuum slot blot manifold. DPCs are detected using antibodies raised against the protein of interest and quantified by normalizing to a DNA loading control. The RADAR assay allows for the detection of specific types of DPCs and the sensitive analysis of the DNA–protein crosslinking activity of various drugs, is adaptable across different cell types and conditions, and requires little specialized equipment.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages14
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • DNA repair
  • DNA-protein crosslink
  • DPC repair
  • RADAR assay
  • Slot blot
  • TOP1-cc
  • TOP2-cc

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


Dive into the research topics of 'Isolation and Immunodetection of Enzymatic DNA–Protein Crosslinks by RADAR Assay'. Together they form a unique fingerprint.

Cite this