ALEXIS Biochemicals: Caspase-2 (active) (human) (recombinant) (His)

ALX-201-135

Revised 15-Oct-08

Caspase-2 (active) (human) (recombinant) (His)
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

Product Specification

MW:	Bands at ~10 and at ~20kDa can be detected by Western blot.
EC:	3.4.22.55
SOURCE/HOST:	Produced in E. coli. Human caspase 2 (aa 18-450) is fused at the C-terminus to a His-tag. When expressed in E. coli, caspase-2 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
CONCENTRATION:	0.2mg/ml
FORMULATION:	Liquid. In 50mM TRIS, pH 8.0, containing 100mM sodium chloride and 50mM imidazole.
ACTIVITY:	The rate of caspase-2 enzymatic hydrolysis 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.
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.

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:

Enzymes • Recombinant Proteins / Fusion Proteins

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