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ALX-201-135 Revised 24-Jan-05
Caspase-2 (active) (human) (recombinant)
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PRODUCT LINE Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY Caspases
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-201-135-C005   5 µg 255.00 USD Add To Cart
Product Specification
EC: 3.4.22.-
SOURCE/HOST: Produced in E. coli.
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 for at least one week when stored at +4°C.
HANDLING: Avoid freeze/thaw cycles.
Product Specific Literature References
Substrate specificities of caspase family proteases: R.V. Talanian, et al.; J. Biol. Chem. 272, 9677 (1997) Abstract; Full Text
General Information
When expressed in E. coli, caspase-2 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
The rate of caspase-2 enzymatic activity can be measured by the release of AFC from the caspase substrate Ac-VDVAD-AFC (Prod. No. ALX-260-112) as emission at 505nm and excitation at 400nm using a spectrofluorometer.
BACKGROUND/TECHNICAL INFORMATION
Enzyme Evaluation Protocol
This protocol is used to measure caspase enzyme activity. The synthetic fluorogenic peptide Ac-VDVAD-AFC (Prod. No. ALX-260-112) is used as the caspase enzyme substrate. The enzyme cleaves the substrate releasing the fluorescent AFC (7-amino-4-trifluoromethylcoumarin). AFC release can be measured by spectrofluorometry using UV.

1. Add 20mM of Ac-VDVAD-AFC (Prod. No. ALX-260-112) 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 AFC liberated from the Ac-VDVAD-AFC using a spectrofluorometer with an excitation wavelength of 400nm and an emission wavelength of 505nm.

Note: Enzyme activity can also be measured using the caspase enzyme substrate Ac-VDVAD-pNA (Prod. No. ALX-260-059) at 37°C in a spectromax at 405nm wavelength.

Swiss-Prot link P42575: Caspase-2 (human) (precursor)
AfCS Signalling Gateway link A000497: Caspase-2 (mouse)
Further Categories Containing This Product:
EnzymesRecombinant Proteins/Fusion Proteins
 
 

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