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 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 cytometry•Immunohistocytochemistry•Paraffi sections •Cryosections

General description

Bromodeoxyuridine (BrdU) is a thymidine analog and is specifically incorporated into DNA during DNA synthesis. Anti-bromodeoxyuridine is used to identify cells that have incorporated BrdU. This immunological detection scheme has several advantages over the use of radioactive thymidine incorporation for identifying cells undergoing replication. Labeling and detection can be performed the same day instead of waiting several days, as required for autoradiography of tritium-labeled cells, and the necessity of using multiple specimens for obtaining the optimal exposure time is eliminated. In addition, antibromodeoxyuridine staining with flow cytometric analysis allows multiple parameters to be evaluated simultaneously.

Monoclonal antibody to 5-bromo-2′-deoxyuridine-5′-monophosphate conjugated to fluorescein.

Immunogen

Apparently, the epitope is inside of 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: 0.5 µg/100 µl (106 cells); Immunohistocytochemistry: 50 µg/mlWorking concentration of conjugate depends on application and substrate. Concentrations should be taken as a guideline.

Specificity

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