title: Detection of apoptotic cells using immunohistochemistry.
abstract: Immunohistochemistry is commonly used to show the presence of apoptotic cells in situ. In this protocol, B-cell lymphoma cells are injected into recipient mice and, on tumor formation, the mice are treated with the apoptosis inducer vorinostat (a histone deacetylase inhibitor). Tumor samples are fixed and sectioned, and fragmented DNA (a feature of apoptotic cells) is end-labeled by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Immunohistochemical methods are then used to detect the labeled DNA and identify B-cell lymphoma cells in the last stage of apoptosis. Because the assay can lead to false-positive results, it is advisable to carry out an additional assay (e.g., immunohistochemistry for active caspase-3) to confirm the presence of apoptotic cells.
authors.affiliation: Gene Regulation Laboratory, Cancer Therapeutics Program, Peter MacCallum Cancer Centre, East Melbourne 3002, Victoria, Australia;
authors.forename: Andrea
authors.initials: A
authors.lastname: Newbold
authors.affiliation: Gene Regulation Laboratory, Cancer Therapeutics Program, Peter MacCallum Cancer Centre, East Melbourne 3002, Victoria, Australia;
authors.forename: Ben P
authors.initials: BP
authors.lastname: Martin
authors.affiliation: Translational Research Laboratory, Cancer Therapeutics Program, Peter MacCallum Cancer Centre, East Melbourne 3002, Victoria, Australia; Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville 3010, Victoria, Australia;
authors.forename: Carleen
authors.initials: C
authors.lastname: Cullinane
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology (LEXOR), Center for Experimental Molecular Medicine, Academic Medical Center, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Michael
authors.initials: M
authors.lastname: Bots
mesh_terms: D000818:Animals; D017209:Apoptosis; D004247:DNA / Q000032:analysis; D053938:DNA Fragmentation; D056572:Histone Deacetylase Inhibitors / Q000008:administration & dosage; D006877:Hydroxamic Acids / Q000008:administration & dosage; D007150:Immunohistochemistry / Q000379:methods*; D020287:In Situ Nick-End Labeling; D016393:Lymphoma, B-Cell / Q000473:pathology*; D051379:Mice; D000077337:Vorinostat
chemical_list: D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D000077337:Vorinostat; D004247:DNA