Immunofluorescent detection of histone 2B on metaphase chromosomes using flow cytometry.

摘要: A monoclonal antibody against histone 2B (anti-H2B) was used as a reagent to stain isolated chromosomes for analysis using flow cytometry. Chromosome suspensions were treated with mouse specific the (clone HBC-7) and then fluorescein-labeled goat anti-mouse-IgM antibody. The also stained DNA content either Hoechst 33258 or propidium iodide. amount of DNA-specific bound each chromosome measured simultaneously order steps in staining protocol is important. Propidium iodide prevents anti-H2B from binding chromosomes, therefore must be added only after labeling completed. In contrast, addition before reduces by 20%–30%. Binding proportional both human Chinese hamster chromosomes. Human bind an average three four times more than do same content. This determined analyzing mixtures results demonstrate that it possible label proteins suspension fluorescent antibodies use these reagents structure