• Product name
    Caspase 9 Assay Kit (Fluorometric)
    See all Caspase 9 kits
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay type
    Enzyme activity
  • Assay time
    2h 00m
  • Product overview

    Abcam's Caspase 9 Assay Kit (Fluorometric) provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEHD. The assay is based on detection of cleavage of substrate LEHD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). LEHD-AFC emits blue light (?max = 400 nm); upon cleavage of the substrate by caspase-9 or related caspases, free AFC emits a yellow-green fluorescence (?max = 505 nm), which can be quantified using a fluorometer or fluorecence microtiter plate reader.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells.


  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    2X Reaction Buffer 4 x 2ml
    Cell Lysis Buffer 1 x 100ml
    DTT 1 x 400µl
    LEHD-AFC (1mM) 1 x 500µl
  • Research areas
  • Relevance
    Caspases are cysteine proteases, expressed as inactive precursors, that mediate apoptosis by proteolysis of specific substrates. Caspases have the ability to cleave after aspartic acid residues. There are two classes of caspases involved in apoptosis; initiators (activation by receptor cluster) and effectors (activation by mitochondrial permeability transition). Proapoptotic signals autocatalytically activate initiator caspases, such as Caspase 8 and Caspase 9. Activated initiator caspases then process effector caspases, such as Caspase 3 and Caspase 7, which in turn cause cell collapse. Caspase 9 (also known as ICE like apoptotic protease 6 (ICE LAP6), apoptotic protease Mch6, and apoptotic protease activating factor 3 (Apaf3)) is a member of the peptidase family C14 that contains a CARD domain. It is active as a heterotetramer and has been reported to have two isoforms. ProCaspase 9 is approximately 47 kD. It is present in the cytosol and, upon activation, translocates to the mitochondria. Caspase 9 is involved in the caspase activation cascade responsible for apoptosis execution and cleaves/activates Caspase 3 and Caspase 6. It becomes activated when recruited to the Apaf1/cytochrome c complex.
  • Alternative names
    • APAF3
    • CASP9
    • Caspase 9
    • ICE LAP6
    • MCH6
    see all

Associated products


  • Caspase 9 activity in Jurkat cells following 24 hour exposure to 50 ng x mL-1 anti-Fas Ab (MBL), 2 uM or 10 uM Camptothecin (CPT; ab120115) with or without 20 nM caspase inhibitor Z-VAD(OMe)-FMK (Zvad; ab120487). Sample input 5-25 uL, background signal subtracted, , duplicates; +/- SD.



This product has been referenced in:
  • Zou ZZ  et al. Synergistic induction of apoptosis by salinomycin and gefitinib through lysosomal and mitochondrial dependent pathway overcomes gefitinib resistance in colorectal cancer. Oncotarget 8:22414-22432 (2017). Functional Studies ; Human . Read more (PubMed: 26461472) »
  • Buvall L  et al. Orellanine specifically targets renal clear cell carcinoma. Oncotarget 8:91085-91098 (2017). Functional Studies . Read more (PubMed: 29207627) »

See all 3 Publications for this product

Customer reviews and Q&As

Thank you for calling Abcam earlier today.

I talked to the lab and they confirmed that you will have to read the signal with the wavelengths for excitation set at 400 nm and emission set at 505 nm. Youonly take one reading.

Hope this ...

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