|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|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10|
|Immunoprecipitation (IP)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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
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