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Cell Biology Apoptosis Intracellular Caspases etc Caspases
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Caspase 9 Assay Kit (Colorimetric) (ab65608)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (1)References (23)

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Functional Studies - Caspase 9 Assay Kit (Colorimetric) (ab65608)

    Key features and details

    • Assay type: Enzyme activity
    • Detection method: Colorimetric
    • Platform: Microplate reader
    • Assay time: 2 hr
    • Sample type: Cell Lysate, Tissue Extracts

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    Overview

    • Product name

      Caspase 9 Assay Kit (Colorimetric)
      See all Caspase-9 kits
    • Detection method

      Colorimetric
    • Sample type

      Tissue Extracts, Cell Lysate
    • Assay type

      Enzyme activity
    • Assay time

      2h 00m
    • Species reactivity

      Reacts with: Mammals
    • Product overview

      Caspase 9 Assay Kit (Colorimetric) ab65608  provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEHD.


      The caspase 9 assay is based on spectrophotometric detection of the chromophore p-nitroanilide (p-NA) after cleavage from the labeled substrate LEHD-p-NA. The p-NA light emission can be quantified using a spectrophotometer or a microtiter plate reader at 400- or 405 nm.

    • Notes

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

      Other caspase and apoptosis assays

      Review the full set of caspase assays, or the apoptosis assay and apoptosis marker guide.

      Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
      It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    • Platform

      Microplate reader

    Properties

    • 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
      Dilution Buffer 1 x 100ml
      DTT 1 x 400µl
      LEHD-pNA (4mM) 1 x 500µl
    • Research areas

      • Cell Biology
      • Apoptosis
      • Intracellular
      • Caspases etc
      • Caspases
      • Kits/ Lysates/ Other
      • Kits
      • Apoptosis Kits
      • Caspase Assays Kits
      • Caspase 9 assay kits
    • 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

    • Related Products

      • Z-LEHD-FMK, caspase-9 inhibitor (ab142026)
      • Q-LEHD-OPh, caspase-9 inhibitor (ab142039)

    Images

    • Functional Studies - Caspase 9 Assay Kit (Colorimetric) (ab65608)
      Functional Studies - Caspase 9 Assay Kit (Colorimetric) (ab65608)Sodhi, Komal et al., Scientific reports?vol. 8,1 9721., Fig 5, doi:10.1038/s41598-018-26768-9
      Caspase-9 activity measured by ab65608. N = 16/group, ∗p < 0.05, ∗∗p < 0.01 vs. corresponding GO concentration groups without pNaKtide.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (23)

    Publishing research using ab65608? Please let us know so that we can cite the reference in this datasheet.

    ab65608 has been referenced in 23 publications.

    • Li C  et al. An investigation on the cytotoxicity and caspase-mediated apoptotic effect of biologically synthesized gold nanoparticles using Cardiospermum halicacabum on AGS gastric carcinoma cells. Int J Nanomedicine 14:951-962 (2019). PubMed: 30787609
    • Ittiudomrak T  et al. a-Mangostin and Apigenin Induced Cell Cycle Arrest and Programmed Cell Death in SKOV-3 Ovarian Cancer Cells. Toxicol Res 35:167-179 (2019). PubMed: 31015899
    • Joshi AU  et al. Drp1/Fis1-mediated mitochondrial fragmentation leads to lysosomal dysfunction in cardiac models of Huntington's disease. J Mol Cell Cardiol 127:125-133 (2019). PubMed: 30550751
    • Wang CH  et al. Dieckol inhibits non-small-cell lung cancer cell proliferation and migration by regulating the PI3K/AKT signaling pathway. J Biochem Mol Toxicol 33:e22346 (2019). PubMed: 31291034
    • Shahbazi B  et al. Cytotoxic effects of Pseudocerastes persicus venom and its HPLC fractions on lung cancer cells. J Venom Anim Toxins Incl Trop Dis 25:e20190009 (2019). PubMed: 31555336
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Question

    Thank you for the information, however i still have question i.e, does the kit contain any buffer for tissue homogenizing or can i use cytosolic tissue homogenate.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 23 2011

    Answer

    Thank you for your enquiry. Indeed, the kits does contain a cell lysis buffer that can be used for tissue homogenising. However, if you have prepared your samples already in your own lysis buffer, you may also dilute 100-200 ug cytosolic protein extract with the lysis buffer included in the kit. Please find attached a more detailed protocol on the assay procedure. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Dec 23 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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