Fluorogenic substrate for caspase-8.
Product Details
Alternative Name: | Caspase-8 substrate (fluorogenic) |
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Sequence: | Ac-Leu-Glu-Thr-Asp-AFC (AFC=7-Amino-4-trifluoromethylcoumarin) |
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Formula: | C31H38F3N5O12 |
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MW: | 729.7 |
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CAS: | 210345-02-1 |
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Purity: | ≥97% (HPLC) |
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Appearance: | White to off-white powder. |
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Solubility: | Soluble in DMSO. |
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Shipping: | Ambient |
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Long Term Storage: | -20°C |
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Use/Stability: | 20mM stock solution in DMSO is stable at -20°C for 6-8 months. When taking out of the freezer wait until it warms up to room temperature, then open it. |
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Handling: | Keep cool and dry. |
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Technical Info/Product Notes: | AFC has an excitation maximum of 400nm and an emission maximum of 505nm. |
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Protocol: | HEPES-Buffer (2x): 40 mM HEPES, pH 7.5, 20% glycerol, 4mM 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: 10µl of substrate stock solution + 1ml 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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Regulatory Status: | RUO - Research Use Only |
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Product Literature References
CRISPR knock out of programmed cell death protein 1 enhances anti-tumor activity of cytotoxic T lymphocytes: Z. Zhao, et al. ; Oncotarget
9, 5208 (2018),
Abstract;
Full Text
Protective effect of 3-hydroxybutyrate against endoplasmic reticulum stress-associated vascular endothelial cell damage induced by low glucose exposure: E. Soejima, et al.; PLoS One
13, e0191147 (2018),
Abstract;
Full Text
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
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;