For assays with recombinant enzymes, esterase should be added: which type of esterase, how much, and can you recommend a specific enzyme?

Name: Z-Val-Ala-Asp (OMe)-FMK Inhibitor
Brand: Novus Biologicals
SKU: NBP2-29392

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.

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

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-29392

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Product Summary
Product Specifications
Preparation & Storage
Background
Product Documents
Specific Notices

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

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.

Product Specifications

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

Formulation, Preparation, and Storage

Formulation

Form: White Solid
Make a stock solution of 20 mM in high purity DMSO (>99.9%).

Concentration

Concentration is not relevant for this product. Please see the protocols for proper use of this product.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: Z-Val-Ala-Asp (OMe)-FMK

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.

Alternate Names

Z Val Ala Asp OMe FMK, Z-Val-Ala-Asp OMe-FMK

Additional Z-Val-Ala-Asp (OMe)-FMK Products

Product Documents for Z-Val-Ala-Asp (OMe)-FMK Inhibitor

Product Datasheets

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Protocols & Manuals

Download NBP2-29392 - Protocol

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COA

Product Specific Notices for Z-Val-Ala-Asp (OMe)-FMK Inhibitor