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Maren Pflueger1, Christoph Wiesner1, Rudolf Lucas2, Josef Atzler3, Michael Katzlinger3, Jim Barry4, Wolfgang Schütt1 and Harald Hundsberger1. 1Medical and Pharmaceutical Biotechnology, Univ. of Applied Sciences, Krems, Austria The caspases are a family of cysteine proteases that are key mediators of apoptosis. There are two types of apoptotic caspases: initiator caspases and effector caspases. Apoptosis can be triggered through either an intrinsic pathway or an extrinsic pathway. Intrinsic apoptosis is initiated through the release of cytochrome C from mitochondria and leads to the activation of initiator caspase 9 and then activation of (effector) caspase 3. In contrast, extrinsic apoptosis is triggered by external death signals such as TNF and CD95 ligand, which activate initiator caspase 8. We describe here a fluoresecence-based assay, which can be performed in either 96- or 384-well cell culture plates. Caspase 3 activation in CD95 ligand treated HepG2 tumor- and immortalized LEC cells can be detected either in the presence or absence of the transcriptional inhibitor actinomycin D. Cell number was normalized using Invitrogen’s CyQUANT* cell proliferation assay with an excitation / emission at 485/530nm, in conjunction with their EnzChek* Caspase-3 Assay, with an excitation/emission profile at 342/441 nm. Detection was performed using the PARADIGM™ Detection Platform from Beckman Coulter. Click here for this poster (PPT) Request More Info on Beckman Coulter Automated Solutions >>
Additional Information
* All trademarks are the property of their respective owners. Where applicable, the PCR process is covered by patents owned by Roche Molecular Systems, Inc., and F. Hoffmann-LaRoche, Ltd. For Research Use Only; not for use in diagnostic procedures. For comments or questions about T3 Update, please contact the T3 Editor. Email this page to a colleague
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