• Non-cytotoxic assay arrests further apoptotic activity via caspase inhibition.
• Cell permeablity permits direct visualization of cytosolic apoptotic events.
• Apoptotic cell population does not diminish over time.
• Add reagent directly to cells. No special buffer or media needed. No preparation of cell lysates required. Simple wash procedure.
• Works in diverse cell lines: human, rodent, Drosophila.
• Can be performed in conjunction with Annexin staining, TUNEL, antibody staining, or with other APO LOGIX reagents on the same population of cells.
• Permits high through-put screening. Protocol can be adapted for ex vivo as well as in situ experiments.
• Yields both quantitative and qualitative results. Gives strong signal with little background noise.
• Mark activity across the range of caspase proteins. Poly-caspase and caspase-specific assays to target caspases 1, 2, 3, 6, 8, 9, or 10 are available.
• Applications - Flow Cytometry, Fluorescence Plate Reader, Fluorescent Microscopy

APO LOGIX Carboxyfluoroscein Caspase Detection Kits label active caspases in living cells undergoing apoptosis. Cell Technology’s probes utilize carboxyfluorescein(FAM)-labeled peptide fluoromethyl ketone (FMK) caspase inhibitors (FAM-peptide-FMK). These FAM-peptide-FMK compounds are both cell permeable and non-cytotoxic during the course of the assay and thus allow the detection of active caspases in living cell systems.

Granzyme B activates procaspase-3 which signals a mitochondrial amplification loop for maximal apoptosis - Sunil S. Metkar, Baikun Wang, Michelle L. Ebbs,Jin H. Kim,Yong J. Lee, Srikumar M. Raja, Christopher J. Froelich - Journal of Cell Biology, Volume 160, Number 6, 875-885, 2003

Small-molecule XIAP inhibitors derepress downstream effector caspases and induce apoptosis of myeloid Leukemia cells - Carter, Gronda, Wang, et.al - Blood, May2005, Vol 105, No 10, pp4043-4050

Fenretinide enhances rituximab-induced cytotoxicity against B-cell lymphoma xenografts through a caspase dependant mechanism - Gopal, Pagel, et.al - Blood, May 2004, Vol 103, No 9, pp 3516-3520.

http://www.nature.com/leu/journal/v19/n1/full/2403560a.html 

Anoxia Is Necessary for Tumor Cell Toxicity Caused by a Low-Oxygen Environment - Papandreou, Krishna, Kaper, et.al - Cancer Research 65, 3171-3178, April 15, 2005

Thimerosal induces DNA Breaks, Caspase 3 Activation, Membrane Damage and Cell Death in Cultured Human Neurons and Fibroblasts - Baskin, Ngo, Didenko - Toxicological Sciences 74, 2003

IFN- Induces Apoptosis in Ovarian Cancer Cells in Vivo and in Vitro - Wall, Burke, Barton, Smyth, Balkwill - Clinical Cancer Research Vol. 9, 2487-2496, July 2003

http://www.nature.com/onc/journal/v24/n24/full/1208542a.html

The removal of extracellular calcium: a novel mechanism underlying the recruitment of N-methyl-d-aspartate (NMDA) receptors in neurotoxicity - Xin, Zhao, Gang, et.al - European Journal of Neuroscience Vol 21, Issue 3, pg 622 - Feb 2005

Deficiency of the Cyclin Kinase inhibitor p21(WAF-1/CIP-1) Promotes Apoptosis of Activated/Memory T Cells, and inhibits spontaneous systemic Autoimmunity - J. Exp Medicine; Vol 199, Number 4, Feb 16 2004; 547-557

Ligand-independent redistribution of Fas (CD95) into lipid rafts mediates clonotypic T cell death - Jagan R Muppidi & Richard M Siegel - Nature Immunology 5, 182 - 189 (2004)

Attenuation of apoptosis in enterocytes by blocking potassium channels - Levitan, Makhina, et. al - Am J Physiol Gastrointest Liver Physiol (July 14, 2005).

Induction of Apoptosis Using Inhibitors of Lysophosphatidic Acid Acyltransferase-ß and Anti-CD20 Monoclonal Antibodies for Treatment of Human Non-Hodgkin's Lymphomas - John M. Pagel, Christian Laugen, Lynn Bonham, Robert C. Hackman, et.al - Clinical Cancer Research Vol. 11, 4857-4866, July 1, 2005. 

Inhibition of PI3K, mTOR and MEK signaling pathways promotes rapid apoptosis in B-lineage ALL in the presence of stromal cell support - Bertrand, Spengeman, Shelton, et.al - Leukemia (2005), 19, 98-102.

Tumor Suppressor DAL-I/4. IB and protein Methylation cooperate in inducing apoptosis MCF-7 breast cancer cells - Molecular Cancer 2006, 5:4

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2. Walker, N. P., R. V. Talanian, K. D. Brady, L. C. Dang, N. J. Bump, C. R. Ferenz, S. Franklin, T. Ghayur, M. C. Hackett and L. D. Hammill. 1994. Crystal Structure of the Cysteine Protease Interleukin-1ß-Converting Enzyme: A (p20/p10)2 Homodimer. Cell 78:343-352.
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5. Kumar, S. 1999. Mechanisms mediating caspase activation in cell death. Cell Death and Differ. 6: 1060-1066.
6. Thornberry, N. A., T. A. Rano, E. P. Peterson, D. M. Rasper, T. Timkey, M. Garcia-Calvo, V. M. Houtszager, P. A. Nordstrom, S. Roy, J. P. Vaillancourt, K. T. Chapman and D. W. Nicholson. 1997. A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J. Biol. Chem. 272(29): 17907-17911.
7. Amstad, P.A., G.L. Johnson, B.W. Lee and S. Dhawan. 2000. An in situ marker for the detection of activated caspases. Biotechnology Laboratory 18: 52-56.
8. Bedner, E., P. Smolewski, P.A. Amstad and Z. Darzynkiewicz. 2000. Activation of caspases measured in situ by binding or fluorochrome-labeled inhibitors of caspases (FLICA): correlation with DNA fragmentation. Exp. Cell Research 259: 308-313.
9. Smolewski, P., E. Bedner, L. Du, T.-C. Hsieh, J. Wu, J. D. Phelps and Z. Darzynkiewicz. 2001. Detection of caspase activation by fluorochrome-labeled inhibitors: multiparameter analysis by laser scanning cytometry. Cytometry 44: 73-82.
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• Lyophilized FAM labeled peptide inhibitor
• 10X Wash Buffer
• 10X Fixative
• Propidium Iodide