Improved bromodeoxyuridine/DNA analysis by anti-BudR monoclonal antibody versus right angle light scatter.
Dynamic cell cycle analysis is based on the incorporation of labelled precursors into DNA. Although antibodies to BrdU are very useful for analysing in flow cells which synthesize DNA, this approach has two main limitations. First, the detection of low incorporating cells is often difficult; second, four parameter flow cytometry is not able to correlate cell cycle to any other cellular marker. We have developed a methodology that, employing an IgGH + L as a second antibody and side scatter instead of propidium iodide fluorescence, allows a better discrimination of BudR+ cells. This approach allows the collection of an extra-fluorescent signal, and the analysis of specific cellular markers within the cell cycle.
Vitale
M
M
Istituto di Anatomia Umana Normale, Università di Bologna, Italy.
Neri
L M
LM
Manzoli
L
L
Galanzi
A
A
Rana
R
R
Antonucci
A
A
Papa
S
S
D000911:Antibodies, Monoclonal; D001973:Bromodeoxyuridine / Q000032:analysis* / Q000276:immunology; D002453:Cell Cycle; D004247:DNA / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods*; D005622:Friend murine leukemia virus; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute; D014407:Tumor Cells, Cultured / Q000166:cytology
D000911:Antibodies, Monoclonal; D004247:DNA; D001973:Bromodeoxyuridine