ALEXIS Biochemicals: Caspase-3 (active) (human) (recombinant)

ALX-201-038

Revised 24-Jan-05

Caspase-3 (active) (human) (recombinant)
PRODUCT LINE	Cell Death / Apoptosis / Autophagy
PRODUCT CATEGORY	Caspases

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-038-C005		5 µg	255.00 USD

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

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

General Information

When expressed in E. coli, caspase-3 spontaneously undergoes autoprocessing to yield the subunits characteristic of the active enzyme.
The rate of caspase-3 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 [1].

BACKGROUND/TECHNICAL INFORMATION

Enzyme Evaluation Protocol
This protocol is used to measure caspase enzyme activity [1]. 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.

Swiss-Prot link P42574: Caspase-3 (human) (precursor)
AfCS Signalling Gateway link A000498: Caspase-3 (mouse)

Further Categories Containing This Product:

Enzymes • Recombinant Proteins / Fusion Proteins

header picture © micronaut.ch, another CS2 website

Online Product Ordering

Clipboard [0]

Your items will be kept only for the duration of your visit.