Overview

  • Product name

    Caspase-3 Immunoassay Kit (Fluorometric)
    See all Cleaved Caspase-3 kits
  • Detection method

    Fluorescent
  • Sample type

    Cell Lysate
  • Assay type

    Quantitative
  • Product overview

    The Caspase 3 Immunoassay Kit (Fluorometric) (ab234045) provides an effective immunosorbent enzyme assay for specific, quantitative detection of caspase 3 activity in microtiter plates. The assay utilizes caspase 3 polyclonal antibody to capture activated caspase 3 from cell lysates. Caspase substrate DEVD-AFC is then added and is cleaved proportionally to the amount of activated caspase 3 in the cell lysate. The cleavage generates free AFC which can be analyzed fluorometrically (Ex/Em = 400 nm/505 nm) using a fluorescence plate reader. The assay ensures absolute specific detection of caspase 3. Other known caspases and non-specific proteases are not detected.

  • Notes

    Activation of caspase 3 plays a key role in initiation of cellular events during apoptosis.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Cell Lysis Buffer 1 x 25ml
    Coating Buffer 1 x 10ml
    Anti-Caspase 3 Antibody (20X) 1 x 0.5ml
    Blocking Buffer 1 x 15ml
    Incubation Buffer 1 x 100ml
    DTT (1M) 1 x 400µl
    DEVD-AFC Substrate (1 mM) 1 x 500µl
    Positive Control (rh-Caspase 3) 1 x 10 units
    Microtite Plate 1 unit
    Adhesive Plate Cover 1 x 2 units
  • Research areas

  • Function

    Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
    -Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. Triggers cell adhesion in sympathetic neurons through RET cleavage.
  • Tissue specificity

    Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system.
  • Sequence similarities

    Belongs to the peptidase C14A family.
  • Post-translational
    modifications

    Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
    S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Alternative names

    • active caspase 3
    • Apopain
    • CASP 3
    • CASP-3
    • CASP3
    • CASP3_HUMAN
    • Caspase 3
    • Caspase-3 subunit p12
    • CPP 32
    • CPP-32
    • CPP32B
    • Cysteine protease CPP32
    • PARP cleavage protease
    • Protein Yama
    • SCA-1
    • SCA1
    • SREBP cleavage activity 1
    • Yama
    see all

Protocols

References

ab234045 has not yet been referenced specifically in any publications.

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