Z-Val-Ala-Asp (OMe)-FMK Inhibitor

Z-Val-Ala-Asp (OMe)-FMK Inhibitor Summary

• Description: This inhibitor is designed as a methyl ester to facilitate cell permeability. If the intended use is on purified or recombinant enzymes, esterase should be added to generate the free carboxyl groups.
• Immunogen: Z-Val-Ala-Asp(OMe)-FMK
  Z-VAD(OMe)-FMK
• Details of Functionality: Mass Spec: M+1=468.2
  Chromatography: TLC:EtOAc:80, Hex: 20, Single Spot, Rf:0.3, Single spot>95%
  H NMR: All functional groups are present
  HPLC: >90%
  Peptide content: 81%
  Formula: C22H30N3O7F
  Molecular Weight: 467

Applications/Dilutions

Publications

Read Publications using NBP2-29392 in the following applications: WB 2 publications

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Form: White Solid
  Make a stock solution of 20 mM in high purity DMSO (>99.9%).

Notes

This inhibitor is designed as a methyl ester to facilitate cell permeability. If the intended use is on purified or recombinant enzymes, esterase should be added to generate the free carboxyl groups.

Alternate Names for Z-Val-Ala-Asp (OMe)-FMK Inhibitor

• Z Val Ala Asp OMe FMK
• Z-Val-Ala-Asp OMe-FMK

Background

Members of the caspase family play key roles in apoptosis and inflammation. Z-VAD(OMe)-FMK is a cell-permeable pan caspase peptide inhibitor that irreversibly binds to the catalytic site of caspases proteases, and inhibits caspasemediated apoptosis by preventing the processing of pro-caspases to their active forms (reviewed in 1-3). The ZVAD( OMe) peptide is O-methylated in the P1 position providing enhanced stability and increased cell permeability. Z-VAD(OMe)-FMK (Z-VAD-FMK) is typically used in assays to inhibit apoptosis. Z-VAD-FMK has been used in many different types of apoptosis assays and published extensively; for example, a PubMed search reveals over 1000 references citing Z-VAD-FMK between 1999 and 2003. Users may want to consult the literature for additional information regarding applications for Z-VAD-FMK.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Inhibitors are guaranteed for 1 year from date of receipt.

Publications for Z-Val-Ala-Asp (OMe)-FMK Inhibitor (NBP2-29392)(6)

Publications using NBP2-29392

• Petris G, Casini A, Sasset L et al. CD4 and BST-2/Tetherin Retro-translocate from ER to Cytosol as Partially Folded and Multimeric Molecules. J Biol Chem 2013-11-20 [PMID: 24257748] (Human)
  
  Details:
  Tetherin transfected HEk293T cells, Fig 6D. Cells were treated with 100 micromolar ZVAD-FMk; Inhibition of PNGase activity.
• Upadhyay S, Liu C, Chatterjee A et al. LKB1/STK11 suppresses cyclooxygenase-2 induction and cellular invasion through PEA3 in lung cancer. Cancer Res. 2006-08-15 [PMID: 16912160] (Human)
  
  Details:
  Products Cited (H1299 human lung cancer cells): 1. Z-VAD-FMK [pan-caspase inhibitor (IMI-2310-1/5], Fig 4C 2. pSuppressorNeo (IMG-800), Fig 7.
• Orlando KA, Pittman RN. Rho kinase regulates phagocytosis, surface expression of GlcNAc, and Golgi fragmentation of apoptotic PC12 cells. Exp Cell Res. 2006-10-15 [PMID: 16904666] (WB)
  
  Details:
  Products cited: 1. ROCK-1 (Cleaved), IMG-383A: WB, Figs 1,2 (serum-starved PC12 cells). IMG-383 detected a 30 kDa cleaved Rock-1 band in serum-starved PC12 cells. The addition of the Broad-Spectrum Caspase Inhibitor Z-VAD-FMK (IMI-2310-1, -5) during serum
• Orlando KA, Stone NL, Pittman RN. Rho kinase regulates fragmentation and phagocytosis of apoptotic cells. Exp Cell Res. 2006-01-01 [PMID: 16259978]
  
  Details:
  Products cited: 1. ROCK-1 (Cleaved), IMG-383A: WB, Fig 3 (tamoxifen-treated Cos7 cells). IMG-383 detected a 30 kDa cleaved Rock-1 band in tamoxifen-treated cells. The addition of the Broad-Spectrum Caspase Inhibitor Z-VAD-FMK (IMI-2310-1, -5) prior to tam
• Xie Q, Lin T, Zhang Y et al. Molecular cloning and characterization of a human AIF-like gene with ability to induce apoptosis. J Biol Chem. 2005-05-20 [PMID: 15764604]
• Caserta TM, Smith AN, Gultice AD et al. Q-VD-OPh, a broad spectrum caspase inhibitor with potent antiapoptotic properties. Apoptosis. 2003-08-01 [PMID: 12815277] (WB)
  
  Details:
  Inhibition of Actinomycin D or TGF beta induced apoptosis in WEHI 231 mouse cells, human Jurkat, or rat HRP-1 (trophoblast) cells: 1. BOC-D-FMK (IMI-2311): Fig 1 (gel analysis of DNA laddering) 2. Z-VAD-FMK (IMI-2310): Fig 1 (gel analysis of DNA laddering) 3. Q-VD-OPH (IMI-2309): Figs 1, 4, 5, 7 (gel analysis of DNA laddering), Figs 2, 3 (TUNEL assay by flow cytometry), Fig 6 (WB analysis of PARP cleavage), Fig 8 (WB analysis of Caspase-3, Caspase-8, and Caspase-9 cleavage) Note: Q-VD-OPH was extensively characterized in this publication and shown to inhibit the major features of apoptosis including caspase activation, DNA laddering, and caspase substrate cleavage.

FAQs for Z-Val-Ala-Asp (OMe)-FMK Inhibitor (NBP2-29392). (Showing 1 - 1 of 1 FAQs).

1. For assays with recombinant enzymes, esterase should be added: which type of esterase, how much, and can you recommend a specific enzyme?
   • When this peptide FMK inhibitor or other OMe peptide inhibitors are used with purified caspases, the ester groups are not rapidly hydrolyzed and the inhibitor-enzyme reaction rate may be reduced. . The ester groups can be removed by pretreating with esterase. For example, dilute the FMK inhibitor stock (or DMSO only for a control) 10-fold into 10 mM borate buffer, pH 8.0, containing 1 unit of esterase (e.g., carboxylic acid ester hydrolase which is available from different vendors) and incubate for 15 minutes on ice. Then dilute an additional 100-fold into the final reaction mixture and incubate again for 15 minutes prior to adding the purified caspase substrate. This is a general protocol, and you may also be able to find other suggestions in the scientific literature.