AntiBrdU

AntiBrdU refers to antibodies that recognize bromodeoxyuridine, a synthetic analog of thymidine used to label newly synthesized DNA in proliferating cells. When BrdU is incorporated into DNA during the S phase, it can later be detected by anti-BrdU antibodies after fixation and DNA denaturation, enabling visualization by immunohistochemistry, immunofluorescence, or flow cytometry. Detection typically requires exposing BrdU within the double helix through denaturation (commonly acid treatment or heat) so that the antibody can bind. Different anti-BrdU antibodies vary in sensitivity and in compatibility with specific denaturation methods, fixation protocols, and tissue types.

Antibodies against BrdU are available as monoclonal and polyclonal reagents from multiple suppliers. Selection depends on experimental context, including tissue type, fixation method, and desired readout. Optimization often includes choosing a suitable denaturation protocol, blocking conditions, and appropriate secondary reagents. Controls commonly used include samples without BrdU labeling and competition with excess unlabeled BrdU to assess specificity.

Applications of anti-BrdU staining include measurement of cell proliferation, determination of S-phase fractions in cell populations, assessment of tissue regenerative or tumor biology, and combination with other markers to define proliferating subpopulations. Limitations include the potential disruption of tissue architecture by denaturation, possible background staining, and the need for BrdU administration to label dividing cells. Alternatives such as EdU-based detection offer a denaturation-free approach, though they require different chemistries and have their own trade-offs.

See also: EdU, thymidine analogs, DNA replication markers.