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Active Motif's DNA Repair Protein ELISAs are DNA-binding ELISAs that detect activity of DNA repair proteins in both mammalian tissue and cell culture extracts. The kits combine a fast, user-friendly format that is more sensitive than gelshift while eliminating the use of radioactivity and providing quantitative results in less than five hours. For more information on how they work, click on the Method tab below.
The GTBP DNA Repair Protein ELISA was designed to study regulation of the G/T Binding Protein (GTBP, also called hMSH6). It contains a 96-well plate to which a double-stranded linear DNA molecule containing a G/T mismatch has been immobilized. GTBP in nuclear extracts binds specifically to this DNA molecule where it is detected and quantitated by addition of GTBP antibody, HRP-conjugated secondary antibody and developing solution. Click the GTBP Info tab below for data and more information.
| Name | Format | Cat. No. | Price | |
|---|---|---|---|---|
| GTBP DNA Repair Protein Kit | 1 x 96 rxns | 51096 | $630 |
 |
| 5 x 96 rxns | 51596 | $2640 |
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| DNA Repair GTBP Manual |
| DNA Repair Profile |
| Cell Biology & Fluorescence Products Brochure |
| MSDS: Sodium Azide |
| MSDS: Sulphuric Acid |
| MSDS: Thimersol |
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).
Applications
GTBP is involved in the maintenance of genomic stability, and therefore represents an excellent pharmacological target for developing drugs to treat cancer. Applications of the GTBP DNA Repair Kit include:
- Profiling of GTBP under various growth and stimulation conditions
- Drug screening and/or potency toward GTBP
- GTBP activity regulation
- Protein structure/function studies of GTBP
Figure 1: Monitoring GTBP binding with the GTBP DNA Repair Kit.
Different amounts of nuclear extracts from unstimulated Raji and Jurkat cells are tested for activity using the GTBP DNA Repair Kit. These curves are provided for demonstration only.
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). Also included are wild-type (DNA lesion cotaining) and mutated oligonucleotides that can be used in competiton assays to verify the specificity of the signal.
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
Figure 1: Flow chart of DNA Repair Protein ELISAs.
DNA repair proteins bind to oligonucleotides that are immobilized in the well. Incubation with primary & secondary antibodies and developing solution enables accurate measurement of DNA repair protein activity using colorimetric readout.
Contents & Storage
One or five 96-well assay plates with plate sealers, primary antibody, secondary antibody, G/T mismatched and non-mismatched oligonucleotides, positive control cell extract, DTT, Protease Inhibitor Cocktail, Lysis, Binding, 10X Washing and 10X Antibody Binding Buffer, and Developing and Stop Solutions. Reagent storage conditions vary from room temperature to -80°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.
