Caspase 9 Inhibitor Drug Detection Kit (ab102497)

Overview

  • Product nameCaspase 9 Inhibitor Drug Detection Kit
    See all Caspase 9 kits
  • Detection methodFluorescent
  • Tests
    100 x 1 assay
  • Sample type
    Adherent cells, Suspension cells
  • Assay typeQuantitative
  • Assay time
    1h 30m
  • Product overview

    Abcam's Caspase 9 Inhibitor Drug Detection Kit provides an effective means for screening caspase inhibitors using fluorometric methods. The assay utilizes synthetic peptide substrate LEHD-AFC (AFC, 7-amino-4-trifluoromethyl coumarin). Active caspase 9 cleaves the synthetic substrate to release free AFC which can then be quantified by fluorometry. Compounds to be screened can directly be added to the reaction and the level of inhibition of caspase 9 activity can be determined by comparison of the fluorescence intensity in samples with and without the testing inhibitors.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Caspases have been shown to play a crucial role in apoptosis induced by various deleterious and physiologic stimuli. Inhibition of caspases can delay apoptosis, implicating a potential role in drug screening efforts.

  • Tested applicationsFunctional Studiesmore details
  • PlatformMicroplate

Properties

  • RelevanceCaspases 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

Applications

Our Abpromise guarantee covers the use of ab102497 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

Caspase 9 Inhibitor Drug Detection Kit images

  • Titration of the caspase 9 inhibitor Z-LEHD-FMK (ab142026) (duplicates; +/- SD).

  • Titration of the caspase inhibitors Z-VAD(OMe)-FMK (ab120487) and Q-LEHD-Oph (ab142039) (duplicates; +/- SD).

  • Titration of caspase inhibitor Z-VAD-FMK (ab102497) (duplicates; +/- SD).

  • Caspase activity (RFU) in presence of 0µM - 40µM of z-VAD-FMK (generic caspase inhibitor), assessed using LEHD-AFC as caspase 9 substrate and following Caspase 9 Inhibitor Drug Detection Kit (ab102493) protocol. Lanes:

    1.- Background Control: no Caspase 9, no z-VMAD-FMK

    2.- Positive Inhibition Control: no Caspase 9, +z-VAD-FMK

    3.- Positive Control: + Caspase 9, no z-VAD-FMK

    4.- Caspase 9 + 2.5µM z-VAD-FMK

    5.- Caspase 9 + 10µM z-VAD-FMK

    6.- Caspase 9 + 40µM z-VAD-FMK

Protocols

References for Caspase 9 Inhibitor Drug Detection Kit (ab102497)

ab102497 has not yet been referenced specifically in any publications.

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1) We have not looked at decreasing volumes for 384-well plates. You may be
able to use these, but it will depend on the amount of caspase in the samples
and hence signal strength can be impacted.
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"