Ac-LETD-AFC

Manuals And Inserts

• Datasheet

Specific 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.

Certificate of Analysis

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