DNA repair proteins act to maintain genome integrity by recognizing, binding to and repairing damaged DNA. Deficiencies in the activity of these proteins is linked to the development of many pathological diseases, including cancer. Understanding and quantifying DNA repair proteins can help elucidate the molecular mechanisms of DNA damage and repair pathways, and understand the damage specificity of a repair protein.

Active Motifs new DNA Repair Kits* are ELISA-based assays for the detection of DNA repair protein activity in mammalian tissue and cell culture extracts. DNA Repair Kits are currently available for studies involving G/T binding protein (GTBP, also called hMSH6) and Ku (Ku70, Ku86), and Replication Protein A (RPA)  RPA70, RPA32.

Active Motifs new DNA Repair Kits offer an improved method for studying DNA repair protein activity. Current techniques used to study DNA damage and repair include EMSAs and Western blots. These methods are time consuming, do not allow for high-throughput and provide only semi-quantitative results. Active Motifs DNA Repair Kits are DNA-binding ELISAs that eliminate radioactivity and provide quantitative results in less than five hours.

The DNA Repair Kit method is simple and efficient and can be used on both tissue and cell culture extracts. Each kit includes a 96-well plate in which multiple copies of a specific double-stranded oligonucleotide have been immobilized. When cellular extract is added, the repair protein of interest binds to the oligonucleotide. Each well is then incubated with a primary antibody that is specific for the repair protein being studied. Addition of a secondary HRP-conjugated antibody and developing solution provides an easily quantified colorimetric readout (Figure 1). This allows you to easily monitor the stimulation or deficiency of DNA repair proteins in a particular cell line, animal model or tumor biopsy.

• Non-radioactive, colorimetric method
• Quantitative results in less than 5 hours
• 10X more sensitive than gelshift
• High-throughput compatible
• Ability to assay both cells and tissues

Figure 1: Flowchart of DNA Repair Pathways