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Anti-Bromodeoxyuridine monoclonal antibody can be used for identifying proliferating cells in blood (1), tissues (2,3), and tumors (4–7), as well as for determining plasma cell labeling indices (8). The antibody may be applied for the qualitative measurement of cell proliferation (BrdU incorporation) on a single-cell level.

Clone
BMC9318

Ig class
mouse IgG₁

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.

Working concentration
Use approximately 0.5 µg Anti-Bromodeoxyuridine-Fluorescein/ 100 µl (10⁶ cells) for flow cytometry and 50 µg/ml Anti-Bromodeoxyuridine-Fluorescein for immunohistochemistry.
These are guidelines only. The optimal concentration should be determined by the investigator. Dilutions should be made in PBS (pH 7.4) containing 0.1% BSA to maintain stability of the antibody.

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.

References

1. Campana, D., Coustan-Smith, E., and Janossy, G. (1988) J. Immunol. Meth. 107: 79.
2. Schutte, B., Reynders, M.M.J., Bosman, F.T., and Blijham, G.H. (1987) J. Histochem. and Cytochem. 35: 1343.
3. Hayashi, Y., Koike, M., Matsutani, M., and Hoshino, T. (1988) J. Histochem. and Cytochem. 36: 511.
4. Hoshino, T., Nagashima, T., Cho, K., Murovic, J., Hodes, J., Wilson, C.,Edwards, M., and Pitts, L. (1986) Int. J. Cancer 38: 369.
5. Nagashima, T., Murovic, J., Hoshino, T., Wilson, C., and Dearmond, S.(1986) J. Neurosurg. 64: 588.
6. Nagashima, T., DeArmond, S., Murovic, J., and Hoshino, T. (1985) Acta Neuropathol. 67: 155.
7. Morstyn, G., Hsu, S.-M., Kinsella, T., Gratzner, H., Russo, A., and Mitchell, J. (1983) J. Clinical Invest. 72: 1844.
8. Greipp, P.R., Witzig, T.E. and Gonchoroff, N.J. (1985) Amer. J. Hematol. 20:289.
9. Vanderlaan, M., Watkins, B., Thomas, C., Dolbeare, F., and Stanker, L. (1986) Cytometry 7: 499.

50 µg Anti-Bromodeoxyuridine-Fluorescein is supplied in 0.5 ml of phosphate-buffered saline (PBS), pH 7.4, containing 0.09% (w/v) sodium azide and 0.2% (w/v) gelatin for stability.

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

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