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| Caspase / Apoptosis
Detection |
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High Throughput Screening
- Apo3HTS |
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Caspase Poly,1,2,3,7,8,9,10
Detection - FAM-FMK |
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Caspase Poly, 1,2,3,8,9,10,
Detection
- (SR-FMK) |
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Antibody Specific Caspase 3 Detection
- Apo Active 3 (FITC) |
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Antibody Specific Caspase 3 Detection
- Apo Active 3 (PE) |
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| NOS/ROS Detection Kits |
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| Fluorescent Enzymatic
Assays |
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| Mitochondria
Membrane Potential detection |
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| aCella - Bioluminescence
Assays Kits |
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| Cytotoxicity |
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| ELISA
Products |
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| Cathepsin B,K,L
Detection Kits |
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Carboxyfluorescein Caspase Detection Kits
™
Apoptosis
detection and in situ labeling of active caspase in live cells.
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| Key Benefits: |
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Non-cytotoxic assay
arrests further apoptotic activity via caspase inhibition.
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Cell permeablity
permits direct visualization of cytosolic apoptotic events.
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Apoptotic cell
population does not diminish over time.
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Add reagent directly
to cells. No special buffer or media needed. No preparation of cell
lysates required. Simple wash procedure.
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Works in diverse cell
lines: human, rodent, Drosophila.
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Can be performed in
conjunction with Annexin staining, TUNEL, antibody staining, or with
other APO LOGIX reagents on the same population of cells.
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Permits high
through-put screening. Protocol can be adapted for ex vivo as well as
in situ experiments.
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Works with
fluorescence microscopy, 96-well fluorescence plate readers, or flow
cytometry.
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Yields both
quantitative and qualitative results. Gives strong signal with little
background noise.
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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.
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Applications -
Flow Cytometry, Fluorescence Plate Reader, Fluorescent Microscope
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| Assay Principle |
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.
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| Citations |
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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| References |
-
Slee, E. A., C.
Adrain, and S. J. Martin. 1999. Serial Killers: ordering caspase
activation events in apoptosis. Cell Death and Differ. 6:1067-1074.
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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.
-
Wilson, K. P., J. F.
Black, J. A. Thomson, E. E. Kim, J. P. Griffith, M. A. Navia, M. A.
Murcko, S. P. Chambers, R. A. Aldape, S. A. Raybuck, and D. J.
Livingston. 1994. Structure and mechanism of interleukin-1 beta
converting enzyme. Nature 370: 270-275.
-
Rotonda, J., D. W.
Nicholson, K. M. Fazil, M. Gallant, Y. Gareau, M. Labelle, E. P.
Peterson, D. M. Rasper, R. Ruel, J. P. Vaillancourt, N. A. Thornberry
and J. W. Becker. 1996. The three-dimensional structure of
apopain/CPP32, a key mediator of apoptosis. Nature Struct. Biol. 3(7):
619-625.
-
Kumar, S. 1999.
Mechanisms mediating caspase activation in cell death. Cell Death and
Differ. 6: 1060-1066.
-
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.
-
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.
-
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.
-
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.
-
Ekert, P. G., J. Silke
and D. L. Vaux. 1999. Caspase inhibitors. Cell Death and Differ.
6:1081-1086.
-
Carcia-Calvo, M., E.
Peterson, B. Leiting, R. Ruel, D. Nicholson and N. Thornberry. 1998.
Inhibition of human caspases by peptide-based and macromolecular
inhibitors. J. Biol. Chem. 273: 32608-32613.
-
Hirata, H., A.
Takahashi, S. Kobayashi, S. Yonehara, H. Sawai, T. Okazaki, K. Yamamoto
and M. Sasada. 1998. Caspases are activated in a branched protease
cascade and control distinct downstream processes in Fas-induced
apoptosis. J. Exp. Med. 187: 587-600.
|
| Kit contents |
- Lyophilized FAM labeled peptide inhibitor
- 10X Wash Buffer
- 10X Fixative
- Propidium Iodide
|
| The
following kits are available: |
Product
|
Catalog No.
|
Size
(No. of Tests) |
Price (US$) |
| FAM-VAD-FMK
Poly caspase detection kit |
FAM100-1 |
25 |
$145 |
FAM100-2
|
100 |
$425 |
FAM-DEVD-FMK
Caspase 3 detection kit
|
FAM200-1 |
25 |
$145 |
FAM200-2
|
100 |
$425 |
| FAM-LETD-FMK
Caspase 8 detection kit |
FAM300-1 |
25 |
$155 |
FAM300-2
|
100 |
$445 |
| FAM-LEHD-FMK
Caspase 9 detection kit |
FAM400-1 |
25 |
$145 |
FAM400-2
|
100 |
$425 |
| FAM-VEID-FMK
Caspase 6 detection kit |
FAM500-1 |
25 |
$195 |
FAM500-2
|
100 |
$495 |
| FAM-YVAD-FMK
Caspase 1 detection kit |
FAM600-1 |
25 |
$155 |
FAM600-2
|
100 |
$445 |
| FAM-VDVAD-FMK
Caspase 2 detection kit |
FAM700-1 |
25 |
$195 |
FAM700-2
|
100 |
$495 |
| FAM-AEVD-FMK
Caspase 10 detection kit |
FAM800-1 |
25 |
$195 |
FAM800-2
|
100 |
$495 |
|
| * Please call 888 7 ASSAYS
(888-727-7297) or email info@celltechnology.com
for volume pricing |
|
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