Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Sabine Pietkiewicz; Clara Wolfe; Jörn H Buchbinder; Inna N Lavrik

doi:10.21769/BioProtoc.2081

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Bio Protoc. 2017 Jan 5;7(1):e2081. doi: 10.21769/BioProtoc.2081.

Authors

Sabine Pietkiewicz^{1

2}, Clara Wolfe¹, Jörn H Buchbinder¹, Inna N Lavrik^{1

3}

Affiliations

¹ Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany.
² Current address: Medical advisory service of social health insurance, Essen, Germany.
³ Federal research center Institute of Cytology and Genetics, Novosibirsk, Russia.

Abstract

Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 and -10 are fully activated by several proteolytic cleavage steps and induce the caspase cascade leading to apoptotic cell death. Analysing the processing of procaspases-8 and -10 by Western blot is a commonly used method to study the induction of apoptosis by death receptor stimulation. To analyse procaspase-8 and -10 cleavage, cells are stimulated with a death ligand for different time intervals, lysed and subjected to Western blot analysis using anti-caspase-8 and anti-caspase-10 antibodies. This allows monitoring the caspase cleavage products and thereby induction of apoptosis.

Keywords: Apoptosis; Caspase-8; Cell death; Cleavage; Proteolysis; Western blot.