Fluorogenic substrate for caspase-8.
|Alternative Name:||Caspase-8 substrate (fluorogenic)|
|Appearance:||White to off-white powder.|
|Solubility:||Soluble in DMSO.|
|Long Term Storage:||-20°C|
|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.|
|Handling:||Keep cool and dry.|
|Technical Info/Product Notes:||AFC has an excitation maximum of 400nm and an emission maximum of 505nm. |
|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.
Reaction mixture without substrate.
Reaction mixture with non-apoptotic cell lysate.
Reaction mixture with apoptotic cell lysate and caspase inhibitor.
Product Literature References
Epidermal growth factor signaling protects from cholestatic liver injury and fibrosis
: J. Svinka, et al.; J. Mol. Med. (Berl.) (2016), 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