Overview

  • Product name
    Z-VAD(OH)-FMK, Irreversible general caspase inhibitor
  • Description
    Irreversible general caspase inhibitor
  • Biological description

    An irreversible general caspase inhibitor. Non cell-permeable and does not require pre-treatment with an esterase. Useful for non-cellular in vitro studies involving isolated, purified or recombinant caspase enzymes. Cell-permeable version also available - Z-VAD (OMe)-FMK ab120487.

  • Purity
    > 94%

Properties

Applications

Our Abpromise guarantee covers the use of ab120382 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 concentration.

Images

  • All lanes : Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436) at 1/1000 dilution

    Lane 1 : Untreated L-929 (Mouse connective tissue fibroblast cells) whole cell lysate
    Lane 2 : L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642), 100 nM Smac mimetic, and 20 µM z-VAD (ab120382) for 8 h and then harvested.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Observed band size: 54 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking and dilution buffer: 5% NFDM/TBST.

  • MLKL (phospho S345) was immunoprecipitated from 1mg of L-929 (Mouse connective tissue fibroblast cells) whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic + 20 µM z-VAD compound (ab120382) for 8h using ab196436 at 1/150 dilution. Western blot was performed from the immunoprecipitate using ab196436 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h;10 µg (Input).
    Lane 2: ab196436 IP in L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab196436 in L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • All lanes : Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436) at 1/1000 dilution

    Lane 1 : L-929 treated with 20 ng/ml TNF alpha (ab9642), 100 nM Smac mimetic, and 20 µM z-VAD (ab120382) for 8 h, whole cell lysate
    Lane 2 : Mouse brain tissue lysate
    Lane 3 : Mouse colon tissue lysate
    Lane 4 : Mouse lung tissue lysate
    Lane 5 : Mouse retina tissue lysate
    Lane 6 : Mouse liver tissue lysate
    Lane 7 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Observed band size: 54 kDa why is the actual band size different from the predicted?


    Exposure time: 50 seconds


    Blocking and diluting buffer: 5% NFDM/TBST.

    MLKL pS345 is a trigger for necroptosis. It is only detectable in infection/cellular damaged (PMID:29229989) or aging tissue (PMID: 28807105) but not in normal tissues.

References

This product has been referenced in:
See all 11 Publications for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE, NOT FOR USE IN HUMANS"

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