 |
| Caspase
/ Apoptosis
Detection |
 |
High
Throughput Screening
- Apo3HTS |
|
Caspase
Poly,1,2,3,7,8,9,10 Detection - FAM-FMK |
|
Caspase
Poly, 1,2,3,8,9,10, Detection
- (SR-FMK) |
|
Antibody
Specific Caspase 3 Detection
- Apo Active 3 (FITC) |
|
Antibody
Specific Caspase 3 Detection
- Apo Active 3 (PE) |
|
| NOS/ROS
Detection Kits |
|
|
| Fluorescent
Enzymatic Assays |
|
|
| Mitochondria
Membrane Potential detection |
|
|
| aCella
- Bioluminescence Assays Kits |
|
|
| Cytotoxicity |
|
|
| ELISA
Products |
|
|
| Cathepsin
B,K,L Detection Kits |
|
|
|
|
|
| |
Carboxyfluorescein
Caspase Detection Kits ™
Apoptosis detection and in situ labeling of active
caspase in live cells.
|
| Key Benefits: |
-
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.
-
Works with fluorescence
microscopy, 96-well fluorescence plate readers,
or flow cytometry.
-
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
Microscope
|
| 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.
|
| 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
|
| 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.
-
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 |
$135 |
FAM100-2
|
100 |
$395 |
FAM-DEVD-FMK Caspase
3 detection kit
|
FAM200-1 |
25 |
$135 |
FAM200-2
|
100 |
$395 |
| 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 |
$135 |
FAM400-2
|
100 |
$395 |
| FAM-VEID-FMK Caspase
6 detection kit |
FAM500-1 |
25 |
$155 |
FAM500-2
|
100 |
$445 |
| 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 |
$155 |
FAM700-2
|
100 |
$445 |
| FAM-AEVD-FMK Caspase
10 detection kit |
FAM800-1 |
25 |
$155 |
FAM800-2
|
100 |
$445 |
|
| * Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for
volume pricing |
|
|
