BPDE-DNA ELISA Kit

Sample

DNA samples such as cell or tissue genomic DNA

Detection Method

iELISA

Intended Use

The BPDE-DNA ELISA Kit is an enzyme immunoassay developed for rapid detection of BPDE-DNA adducts. The quantity of BPDE adduct in DNA samples is determined by relative comparison of a known BPDE-DNA standard curve. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown protein samples.

Contents of Kit

Box 1
1. DNA High-Binding Plate: One 96-well strip plate.
2. DNA Binding Solution: One 6 mL bottle.
3. Anti-BPDE Antibody (1000X): One 20 μL vial of anti-BPDE-I antibody.
4. Secondary Antibody, HRP Conjugate (1000X): One 50 μL vial.
5. Assay Diluent: One 50 mL bottle.
6. 10X Wash Buffer: One 100 mL bottle.
7. Substrate Solution: One 12 mL amber bottle.
8. Stop Solution: One 12 mL bottle.
9. Reduced DNA Standard: One 200 μL vial of 0.2 mg/mL reduced DNA in TE Buffer
Box 2
1. BPDE-DNA Standard: One 30 μL vial of 0.1 mg/mL BPDE-DNA in TE Buffer.

Storage

Upon receipt, aliquot and store the Reduced DNA and BPDE-DNA Standards at -20°C to avoid multiple freeze/thaw cycles. Store all other kit components at 4°C.

General Description

Polycyclic aromatic hydrocarbons (PAHs) are potent, ubiquitous atmospheric pollutants commonly associated with oil, coal, cigarette smoke, and automobile exhaust fumes. Some PAH compounds are also found in cooked foods (e.g. grilled meat, smoked fish) and have been identified as mutagenic and carcinogenic. The toxicity of some PAHs has been demonstrated to induce malignant tumors in animal models and is also commonly believed to significantly contribute to human cancers.

One PAH compound, benzo(a)pyrene, is notable for being the first chemical carcinogen to be discovered. Benzo(a)pyrene is a five-ring PAH known to be a procarcinogen; its mechanism of carcinogenesis is dependent on a 3-step enzymatic metabolism (Fig. 1 below) to the final mutagen benzo(a)pyrene diol epoxide (BPDE). Very reactive, BPDE binds covalently to proteins, lipids, and DNA (guanine residues) to produce BPDE adducts. If left unrepaired, DNA adducts may lead to permanent mutations resulting in cell transformation and ultimately tumor development.

Figure 1: Benzo(a)pyrene catalyzed to various metabolites by Cytochrome P450 enzymes (CYP) and epoxide hydrolase (EH), resulting in the final carcinogen BPDE.

General Notes

The BPDE-DNA ELISA Kit is an enzyme immunoassay developed for rapid detection of BPDE-DNA adducts. The quantity of BPDE adduct in DNA samples is determined by relative comparison of a known BPDE-DNA standard curve. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown protein samples.