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Ac-LETD-AFC

Caspase-8 substrate
 
ALX-260-118-M005 5 mg 194.00 USD
 
ALX-260-118-M010 10 mg 366.00 USD
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Fluorogenic substrate for caspase-8.

Product Specification

Alternative Name:Caspase-8 substrate (fluorogenic)
 
Sequence:Ac-Leu-Glu-Thr-Asp-AFC (AFC=7-Amino-4-trifluoromethylcoumarin)
 
Formula:C31H38F3N5O12
 
MW:729.7
 
CAS:210345-02-1
 
Purity:≥97% (HPLC)
 
Appearance:White to off-white powder.
 
Solubility:Soluble in DMSO.
 
Shipping:Ambient
 
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.

Suggested controls:
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;

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