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| Caspase
/ Apoptosis
Detection |
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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) |
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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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APO
LOGIX Sulforhodamine Caspase Detection Kit ™
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.
-
Applications - Works with fluorescence
microscope, 96-well fluorescence plate readers
-
Yields both quantitative and qualitative
results. Gives strong signal with
little background noise.
|
| Assay Principle |
APO LOGIX
SR kits contain a generic sulforhodamine labeled
caspase inhibitor (sulforhodamine-peptide-fluoromethyl
ketone). This reagent is cell permeable and is used
on whole cells to detect apoptosis. Apoptotic cells
are detected by a fluorescence plate reader or fluorescence
microscope using an excitation source at 550nm and
measuring emission at 595nm. The assay takes about
1 hr to complete
|
| 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 SRily 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 SR-VAD-FMK
- 10X Wash Buffer
- 10X fixative
|
| The following
kits are available: |
Product
|
Catalog No.
|
Size
(No. of Tests) |
Price (US$) |
| SR-VAD-FMK Poly
caspase detection kit |
SR100-1 |
25 |
$195 |
SR100-2
|
100 |
$525 |
SR-DEVD-FMK Caspase
3 detection kit
|
SR200-1 |
25 |
$195 |
SR200-2
|
100 |
$525 |
|
| * Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for
volume pricing |
|
|
