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Caspase-7 (human), (recombinant) (active)

 
ALX-201-040-C005 5 µg 196.00 USD
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Product Specification

Alternative Name:Mch3, CMH-1
 
Source:Produced in E. coli.
 
EC:3.4.22.-
 
UniProt ID:P55210
 
Formulation:Liquid. 5µg in 25µl of 50mM TRIS (pH 8.0) containing 100mM sodium chloride and 50mM imidazole.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Use/Stability:Stable at least one week when stored at +4°C.
 
Handling:Avoid freeze/thaw cycles.
 
Scientific Background:When expressed in E. coli, caspase-7 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
The rate of caspase-7 enzymatic hydrolysis can be measured by the release of AMC from the caspase substrate Ac-DEVD-AMC (Prod. No. ALX-260-031) as emission at 440nm and excitation at 380nm using a spectrofluorometer.
 
Protocol:Enzyme Evaluation Protocol
This protocol is used to measure caspase enzyme activity. The synthetic fluorogenic peptide Ac-DEVD-AMC (Prod. No. ALX-260-031) is used as the caspase enzyme substrate. The enzyme cleaves the substrate releasing the fluorescent AMC (7-amino-4-methylcoumarin). AMC release can be measured by spectrofluorometry using UV.

1. Add 20mM of Ac-DEVD-AMC (Prod. No. ALX-260-031) to 1ml of assay buffer (20mM PIPES, 100mM sodium chloride, 10mM DTT, 1mM EDTA, 0.1% (w/v) CHAPS, 10% sucrose, pH 7.2). Add the appropriate amount of active caspase to the mixture (50ng/ml).
2. Incubate for 1 hour at 37°C.
3. Measure the AMC liberated from the Ac-DEVD-AMC using a spectrofluorometer with an excitation wavelength of 380nm and an emission wavelength of 440nm.

Note: Enzyme activity can also be measured using the caspase enzyme substrate Ac-DEVD-pNA (Prod. No. ALX-260-033) at 37°C in a spectromax at 405nm wavelength.
 

Product Literature References

Biochemical characteristics of caspases-3, -6, -7, and -8: H.R. Stennicke & G.S. Salvesen; J. Biol. Chem. 272, 25719 (1997), Abstract; Full Text

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For Research Use Only. Not for use in diagnostic procedures.
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