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The G/T binding protein (GTBP, also called MSH6) is a mismatch repair protein that binds as a heterodimer to DNA. GTBP specifically recognizes mismatched G/T base pairs in double-stranded DNA where it triggers excision and repair. Deficiencies in the repair of mismatched G/T base pairs can lead to microsatellite instability and is also linked to hereditary nonpolyposis colon cancer (HNPCC).
DNA repair proteins act to maintain genome integrity by recognizing, binding to and repairing damaged DNA. Deficiencies in the activity of these proteins are 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.
The DNA Repair Protein ELISA Kit advantage
DNA Repair Protein ELISAs offer an improved method for studying DNA repair protein activity. Current techniques used to study DNA damage and repair include EMSAs and Western blots, which are time consuming, do not allow for high-throughput and provide only semi-quantitative results. In contrast, Active Motif’s DNA Repair Kits are DNA-binding ELISAs that eliminate radioactivity and provide quantitative results in less than five hours.
Each kit includes a 96-well plate in which multiple copies of a specific damaged oligonucleotide have been immobilized. When cellular extract is added, the repair protein of interest binds to the damaged DNA. 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).
Key features
- Non-radioactive, colorimetric method
- Quantitative results in less than 5 hours
- 10X more sensitive than gelshift
- High and low-throughput compatible – 96-stripwell format
- Ability to assay both cells and tissues
