Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)
Key features and details
- Irreversible general caspase inhibitor
- CAS Number: 220644-02-0
- Purity: > 94%
- Soluble in DMSO to 20 mM
- Form / State: Solid
- Source: Synthetic
Overview
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Product name
Z-VAD(OH)-FMK, Irreversible general caspase inhibitor -
Description
Irreversible general caspase inhibitor -
Purity
> 94% -
CAS Number
220644-02-0 -
Chemical structure
Properties
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Molecular weight
453.46 -
Molecular formula
C21H28FN3O7 -
Sequence
VAD (Modifications: N-terminal benzyloxycarbonyl; C-terminal FMK) -
PubChem identifier
5497170 -
Storage instructions
Store at -20°C. Store under desiccating conditions. The product can be stored for up to 12 months. -
Solubility overview
Soluble in DMSO to 20 mM -
Handling
Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one week. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.
Need more advice on solubility, usage and handling? Please visit our frequently asked questions (FAQ) page for more details.
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SMILES
FCC(=O)C(CC(=O)O)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)OCc1ccccc1)C(C)C -
Source
Synthetic
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Research areas
Images
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Western blot - Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)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 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Functional Studies - Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382) Davis J et al. PLoS One. 2013; 8(12): e82053. doi: 10.1371/journal.pone.0082053 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/ZVAD-fmk treatment does not correct muscular dystrophy in Nol3-/-Sgcd-/- mice.
A: Muscle weights normalized to tibial length of quadriceps of mice treated with or without ZVAD-fmk for 4 weeks
B: Creatine Kinase levels
C: Quantification of the time to exhaustion as assessed by involuntary treadmill running measured from vehicle or ZVAD-fmk treated WT and Nol3-/- Sgcd-/- mice
D: Histologic images taken at 100x of Masson’s trichrome stained sections of quadriceps from vehicle or ZVAD-fmk treated WT and Nol3-/-Sgcd-/- mice
Davis J et al. PLoS One. 2013; 8(12): e82053. doi: 10.1371/journal.pone.0082053
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Immunoprecipitation - Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)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.
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Western blot - Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)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 secondsBlocking 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.