|Flow Cytometry (Flow)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10|
|Immunoprecipitation (IP)||Assay Dependent|
|Tested Species reactivity||Chemical|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||BPDE-I-DNA complexed with methylated BSA|
|Storage buffer||PBS with 0.1% BSA|
|Contains||0.02% sodium azide|
|Storage conditions||4° C|
This antibody recognizes BPDE-I-DNA (PAH-DNA).
Flow Cytometry: samples were fixed in 2% paraformaldehyde and permeabilized with 0.2% triton X-100/0.1% sodium citrate. Samples were treated with protK and RNase. To denature DNA, samples were incubated with 4n HCl. Samples were blocked with 5% normal serum before incubation with the monoclonal antibody.
ELISA: plates were coated with 50 ng/well BPDE-DNA in 50mM Tris-buffer pH7.5 o/n at 4°C. Plates were blocked 1% FCS. DNA samples, 4µg, were mixed with 5D11 and added to the well. Detection was performed using a Goat anti-Mouse IgG (AP) for 90 min. at 37°C.
IHC (P): 5 µm sections were RNase and prot-K treated. DNA was denatured with 4N HCl and neutralized with 50mM Tris base. Sections were blocked with 1.5% normal horse serum prior to staining.
A number of chemicals, including polycyclic aromatic hydrocarbons (PAHs), have been shown to bind to DNA. This DNA damage can occur both early and late in the malignant process, thereby acting as an initiator and assisting in the progression of tumors. PAHs are released into the environment following incomplete combustion of organic materials. The most common sources of PAHs are from smoking and from consuming broiled or grilled foods. Human exposure to PAHs comes from various occupational, environmental, dietary and medicinal sources. Benzo[a]pyrene is a representative PAH. Antibodies to benzo[a]pyrenediol-epoxide modified DNA (BPDE-DNA) can be used to identify polycyclic aromatic hydrocarbon (PAH)-DNA adducts. Exposure to this group of compounds is believed to be carcinogenic.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein Aliases: Benzo[a]pyrenediol-epoxide; BPDE-DNA; BPDE-I-DNA; PAH-DNA
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