Apoptosis-associated caspase activation assays.
Caspases are aspartate-directed cysteine proteases that cleave a diverse group of intracellular substrates to contribute to various manifestations of apoptosis. These proteases are synthesized as inactive precursors and are activated as a consequence of signaling induced by a wide range of physiological and pathological stimuli. Caspase activation can be detected by measurement of catalytic activity, immunoblotting for cleavage of their substrates, immunolabeling using conformation-sensitive antibodies or affinity labeling followed by flow cytometry or ligand blotting. Here we describe methods for each of these assays, identify recent improvements in these assays and outline the strengths and limitations of each approach.
Kaufmann
Scott H
SH
Division of Oncology Research, Mayo Clinic College of Medicine, 200 First Street, S.W., Rochester, MN 55905, USA. Kaufmann.Scott@Mayo.edu
Lee
Sun-Hee
SH
Meng
X Wei
XW
Loegering
David A
DA
Kottke
Timothy J
TJ
Henzing
Alexander J
AJ
Ruchaud
Sandrine
S
Samejima
Kumiko
K
Earnshaw
William C
WC
D000345:Affinity Labels; D000818:Animals; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism*; D002645:Chickens; D004789:Enzyme Activation; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting
D000345:Affinity Labels; D020169:Caspases