Caspase fluorometric substrate set

Manuals And Inserts

• Manual
• Datasheet

Specific Protocol

Assay Procedure:
1. Induce apoptosis in cells by desired method. Concurrently incubate a control culture without induction.
2. Count cells and pellet 1-5x10⁶ cells or use 50-200µg cell lysates if protein concentration has been measured.
3. Resuspend cells in 50µl of chilled Cell Lysis Buffer.
4. Incubate cells on ice for 10 minutes.
5. Add 50µl of 2x Reaction Buffer containing 10mM DTT to each sample.
6. Add 5µl of the 1mM AFC conjugated substrates (50µM final conc.) into each tube individually and incubate at 37°C for 1-2 hours.
7. Read samples in a fluorometer equipped with a 400-nm excitation filter and a 505-nm emission filter. For a plate-reading set-up, transfer the samples to a 96-well plate. You may perform the entire assay directly in a 96 well plate.
Fold-increase in caspase activity can be determined by comparing these results with the level of the uninduced control.
Note: Background reading from cell lysates and buffers should be subtracted from the readings of both induced and the uninduced samples before calculating fold increase in caspase activity.

Certificate of Analysis

Please enter the lot number as featured on the product label

Lot No. :

SDS

Enzo Life Science provides GHS Compliant SDS

• English (EU) SDS

If your language is not available please fill out the SDS request form