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Caspase-3 substrate
Fluorogenic substrate for caspase-3 (CPP32), with a Km=9.7µM) and related cysteine proteases. Sequence is based on PARP cleavage at Asp216 for caspase-3. Similar to Ac-DEVD-AMC (Prod. No. ALX-260-031) but the AFC fluorophore has a greater Stokes’ shift upon cleavage than AMC. Reaction can be monitored quantitatively or visually using a hand-held long-UV lamp and visualizing a blue to green shift in fluorescence upon cleavage. Ex.: 400nm, Em.: 505nm.
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Product Details
| Alternative Name |
Caspase-3 substrate (fluorogenic) |
|---|---|
| Appearance |
White to off-white powder. |
| CAS |
201608-14-2 |
| Formula |
C30H34F3N5O13 |
| MW |
729.6 |
| Peptide Content |
65-95% |
| Purity |
≥96% (HPLC) |
| Sequence |
Ac-Asp-Glu-Val-Asp-AFC (AFC = 7-Amino-4-trifluoromethylcoumarin) |
| Solubility |
Soluble in dimethyl formamide, DMSO or methanol; slightly soluble in water (0.4mg/ml). |
| Technical Info / Product Notes |
AFC has an excitation maximum of 400nm and an emission maximum of 505nm. |
Handling & Storage
| Use/Stability |
As indicated on product label or CoA when stored as recommended. |
|---|---|
| Handling |
Protect from light. Keep cool and dry. |
| Long Term Storage |
-20°C |
| Shipping |
Ambient Temperature |
| Regulatory Status |
RUO – Research Use Only |
|---|
- SUMOylation controls Hu antigen R posttranscriptional activity in liver cancer: S. Lachiondo-Ortega, et al.; Cell Rep. 43, 113924 (2024), Abstract
- Colitis ameliorates cholestatic liver disease via suppression of bile acid synthesis: Gui, W., Hole, M. J., et al.; Nat. Commun. 14, 3304 (2023), Abstract
- Helicobacter pylori outer membrane vesicles induce astrocyte reactivity through nuclear factor-κappa B activation and cause neuronal damage in vivo in a murine model: E. Palacios, et al.; J. Neuroinflammation 20, 66 (2023), Abstract
- CARD-only proteins regulate in vivo inflammasome responses and ameliorate gout: S. Devi, et al.; Cell Rep. 42, 112265 (2023), Abstract
- Antitumoral Activity of Leptocarpha rivularis Flower Extracts against Gastric Cancer Cells: N. Carrasco, et al.; Int. J. Mol. Sci. 24, 1439 (2023), Abstract
- Sublethal cytochrome c release generates drug-tolerant persister cells: Kalkavan, H., Chen, M. J., et al.; Cell 185, 3356 (2022), Abstract
- Characterization of Anti-Cancer Activities of Violacein: Actions on Tumor Cells and the Tumor Microenvironment: Dahlem, C., Chanda, S., et al.; Front. Oncol. 12, 872223 (2022), Abstract
- Autophagy displays divergent roles during intermittent amino acid starvation and toxic stress-induced senescence in cultured skeletal muscle cells: D. Bloemberg & J. Quadilatero; J. Cell. Physiol. 236, 3099 (2021), Application(s): Caspase-3 activity assay on cell lysates, Abstract
- Ibrutinib blocks YAP1 activation and reverses BRAFi resistance in melanoma cells: Misek, S., Newbury, P., et al.; bioRxiv , (2020)
- Delineation of cell death mechanisms induced by synergistic effects of statins and erlotinib in non-small cell lung cancer cell (NSCLC) lines: Otahal, A., Aydemir, D., et al.; Sci. Rep. 10, 959 (2020), Abstract
- Toll-Like Receptor 2 Release by Macrophages: An Anti-inflammatory Program Induced by Glucocorticoids and Lipopolysaccharide: J. Hoppstadter, et al.; Front. Immunol. 10, 1634 (2019), Abstract — Full Text
- Additive polyplexes to undertake siRNA therapy against CDC20 and survivin in breast cancer cells: M.B. Parmar, et al.; Biomacromolecules 19, 4193 (2018), Abstract
- Sub-lethal oxidative stress induces lysosome biogenesis via a lysosomal membrane permeabilization-cathepsin-caspase 3-transcription factor EB-dependent pathway: S.M. Leow, et al.; Oncotarget 8, 16170 (2017), Abstract — Full Text
- Nimbolide reduces CD44 positive cell population and induces mitochondrial apoptosis in pancreatic cancer cells: S. Kumar, et al.; Cancer Lett. 413, 82 (2017), Abstract
- Epidermal growth factor signaling protects from cholestatic liver injury and fibrosis: J. Svinka, et al.; J. Mol. Med. (Berl.) 95, 109 (2017), Application(s): Detection of active caspases, liver hepatocytes, Abstract — Full Text
- Decreased Poly(ADP-Ribose) Polymerase 1 Expression Attenuates Glucose Oxidase-Induced Damage in Rat Cochlear Marginal Strial Cells: Y. Zhang, et al.; Mol. Neurobiol. 53, 5971 (2016), Abstract
- Mechanism of neem limonoids-induced cell death in cancer: Role of oxidative phosphorylation: N. Yadav, et al.; Free Radic. Biol. Med. 90, 261 (2016), Application(s): Quantification of apoptosis and caspase activity measurement, Abstract
- Artesunate induces ROS-dependent apoptosis via a Bax-mediated intrinsic pathway in Huh-7 and Hep3B cells: Y. Pang, et al.; Exp. Cell Res. 16, 30161 (2016), Application(s): Fluorometric assay for caspase-3 activity, Abstract
- Organ specific alteration in caspase expression and STK3 proteolysis during the aging process: M. Lessard-Beaudoin, et al.; Neurobiol. Aging 47, 50 (2016), Application(s): Caspase activity assays, Abstract — Full Text
- Post-transcriptional control of executioner caspases by RNA-binding proteins: D. Subasic, et al.; Genes Dev. 30, 2213 (2016), Application(s): Caspase activity reporter, HeLa Kyoto cells, Abstract — Full Text
- The C-terminal domains of apoptotic BH3-only proteins mediate their insertion into distinct biological membranes: V. Andreu-Fernandez, et al.; J. Biol. Chem. 291, 25207 (2016), Application(s): Cell culture to monitor caspase activity in cell extracts, Abstract — Full Text
- Involvement of Bim in Photofrin-Mediated Photodynamically Induced Apoptosis: X. Wang, et al.; Cell Physiol. Biochem. 35, 1527 (2015), Application(s): Cell Culture , Abstract — Full Text
- Elevation of soluble guanylate cyclase suppresses proliferation and survival of human breast cancer cells: H. C. Wen, et al.; PLoS One 10, e0125518 (2015), Application(s): Cell Culture, Caspase 3 activity assay, Abstract — Full Text
- Redox regulation of metabolic and signaling pathways by thioredoxin and GLUTAREDOXIn in nos-3 overexpressing hepatoblastoma cells: R. González, et al.; Redox Biol. 6, 122 (2015), Application(s): Cell Culture, Fluorescence, Abstract
- Bothropoides pauloensis venom effects on isolated perfused kidney and cultured renal tubular epithelial cells: A.D. Marinho, et al.; Toxicon 108, 126 (2015), Application(s): Caspase activity in rat isolated kidney, Abstract
- Oxidative phosphorylation-dependent regulation of cancer cell apoptosis in response to anticancer agents: N. Yadav, et al.; Cell Death Dis. 5, e1969 (2015), Application(s): Cell culture, Fluorescence, Abstract — Full Text
- A novel TNFR1-triggered apoptosis pathway mediated by class IA PI3Ks in neutrophils: B. Geering, et al.; Blood 117, 5953 (2011), Application(s): Caspase activity detected in human neutophils, Abstract — Full Text
- Different subcellular distribution of caspase-3 and caspase-7 following Fas-induced apoptosis in mouse liver: J.M. Chandler, et al.; J. Biol. Chem. 273, 10815 (1998), Abstract — Full Text
- Caspase-7 is activated during lovastatin-induced apoptosis of the prostate cancer cell line LNCaP: M. Marcelli, et al.; Cancer Res. 58, 76 (1998), Abstract
- BAX-induced cell death may not require interleukin 1 β-converting enzyme-like proteases: J. Xiang, et al.; Proc. Natl. Acad. Sci. USA 93, 14559 (1996), Abstract
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Last modified: May 29, 2024
Datasheet, Manuals, SDS & CofA
Manuals And Inserts
Specific Protocol
HEPES-Buffer (2x): 40 mM HEPES, pH 7.5, 20% glycerol, 4 mM DTT. Dilute to 1x with sterile distilled water prior to use.
Substrate: Prepare 20 mM stock solution in DMSO
- Induce apoptosis and prepare cell lysate or use recombinant caspase.
- Prepare reaction buffer: 10 µl of substrate stock solution + 1 ml 1x HEPES-Buffer for each reaction.
- Add an appropriate amount of cell lysate
(50-100 µl; should be titrated) or recombinant caspase to reaction buffer.
- Incubate for 1 hour at 37 °C.
- Measure with spectrofluorometer: 400 nm excitation wavelength, 505 nm emission wavelength.
- Suggested controls:
- Reaction mixture without substrate.
- Reaction mixture with non-apoptotic cell lysate.
- Reaction mixture with apoptotic cell lysate and caspase inhibitor.
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