Analysis Note

Anti-Bromodeoxyuridine shows 10% cross reaction with iodo-deoxyuridine, but no cross reaction to fluoro-deoxyuridine.No cross reaction to any endogenous thymidine or uridine.Cross reactivity with 5-Br-UTP has not been tested but it is suggested that there is a good chance for reaction, because the only difference is an absent hydroxyl group on the ribose distal to the bromine substitution.

Application

Anti-bromodeoxyuridine (Anti-BrdU) antibody is suitable for monitoring proliferating cells in blood, tissues, and tumors, as well as for determining BrdU incorporation on a single-cell level using: Flow cytometryImmunohistocytochemistryCryosectionsParaffin sections

General description

Anti-Bromodeoxyuridine is a monoclonal antibody to 5-bromo-2′-deoxyuridine-5′-monophosphate.

Immunogen

The epitope is apparently inside the DNA helix. DNA has to be denatured to ssDNA before antibody efficiently binds to DNA-BrdU.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Physical form

Solution, stabilized with phosphate buffered saline, pH 7.4, containing 0.09% (w/v) sodium azide and 0.2% (w/v) gelatin.

Preparation Note

Working concentration: Flow cytometry: 2 µg/ml (0.2 µg/100 µl/106 cells); Immunohistocytochemistry: 6 µg/mlWorking concentration of conjugate depends on application and substrate. Dilutions should be made in PBS (pH 7.4) containing 0.1% BSA to maintain stability of the antibody.

Quality

The antibody is ≥90% pure as determined by SDS-PAGE with Coomassie-blue staining, and by HPLC.

Specifications

Preparation: BALB/c mice were immunized with a bromodeoxyuridine-bovine serum albumin conjugate. Lymphocytes isolated from the spleen were fused with Ag8.653 myeloma cells to create the BMC 9318 clone. The antibody was produced in ascites in BALB/c mice and purified by ion-exchange chromatography.No. of tests: 250 (Flow cytometry)

Specificity

The antibody specifically binds to bromodeoxyuridine and crossreacts with iodouridine (10%). Anti-bromo-deoxyuridine does not crossreact with fluorodeoxy-uridine, nor with any endogenous cellular components such as thymidine or uridine.