title: Detecting caspase activity in Drosophila larval imaginal discs.
abstract: Caspases are a highly specialized class of cell death proteases. Since they are synthesized as inactive full-length zymogens, activation--at least of effector caspases and to some extent also of initiator caspases-requires a proteolytic cleavage event, generating a large and a small subunit, two of each forming the active caspase. The proteolytic cleavage event generates neo-epitopes at both the C-terminus of the large subunit and the N-terminus of the small subunit. The cleaved Caspase-3 (CC3) antibody was raised against the neo-epitope of the large subunit and thus detects only cleaved, but not full-length, Caspase-3. Although raised against human cleaved Caspase-3, the CC3 antibody cross-reacts in other species and detects cleaved caspases, most notably DrICE and Dcp-1, in Drosophila. This protocol describes the procedure for use of the CC3 antibody to detect caspase activity in larval imaginal discs in Drosophila.
authors.affiliation: Department of Cancer Biology, University of Massachusetts Medical School, Worcester, MA, USA.
authors.forename: Caitlin E
authors.initials: CE
authors.lastname: Fogarty
authors.forename: Andreas
authors.initials: A
authors.lastname: Bergmann
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D053148:Caspase 3 / Q000235:genetics / Q000302:isolation & purification*; D004331:Drosophila melanogaster / Q000201:enzymology; D006801:Humans; D060227:Imaginal Discs / Q000201:enzymology; D007814:Larva / Q000201:enzymology; D008967:Molecular Biology / Q000379:methods*
chemical_list: D053148:Caspase 3