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Caspase-3 (rat), (recombinant)

ALX-201-078-C005 5 µg 345.00 USD
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Contains both the proenzyme and the processed (active) form.

Product Specification

Alternative Name:CPP32, Yama, Apopain
Source:Produced in E. coli.
UniProt ID:P55213
Formulation:Liquid. 5µg of caspase-3 in 50mM HEPES, pH 7.4, 100mM NaCI, 0.5% CHAPS, 10mM DTT, 1mM EDTA and 50% glycerol.
Purity:≥95% (SDS-PAGE)
Specific Activity:≥500U/µg protein. One unit is defined as the amount of enzyme that cleaves 1nmol of the caspase-substrate Z-DEVD-AMC per hour at 20°C in a reaction solution containing 20µM substrate, 50mM HEPES, pH 7.4, 100mM NaCl, 0.5% CHAPS, 10mM DTT, 1mM EDTA and 10% glycerol.
Application Notes:In combination with caspase-3 activity assays, this product is useful in biological screening of caspase substrates and inhibitors. It can also be used as a positive control in caspase-3 assays as well as a positive control  in Western blot.
Shipping:Shipped on Dry Ice
Short Term Storage:-20°C
Long Term Storage:-80°C
Use/Stability:Stability at -20°C: 1-2 weeks
Handling:Avoid freeze/thaw cycles.
Protocol:Enzyme Evaluation Protocol (general)
This protocol can be used to measure caspase enzyme activity. A synthetic fluorogenic peptide, e. g. Ac-DEVD-AMC (Prod. No. ALX-260-031) is used as the caspase enzyme substrate. The enzyme cleaves the substrate releasing the fluorescent AMC. AMC release can be measured by spectrofluorometry using UV.

1. Add 20µM of Ac-DEVD-AMC to 1 ml of assay buffer (20mM PIPES, pH 7.2, 100mM sodium chloride, 10mM DTT, 1mM EDTA, 0.1% (w/v) CHAPS, 10% sucrose). Add the appropriate amount of active caspase to the mixture (e.g. 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.

Product Literature References

Differential expression of apoptotic protease-activating factor-1 and caspase-3 genes and susceptibility to apoptosis during brain development and after traumatic brain injury: A.G. Yakovlev, et al.; J. Neurosci. 21, 7439 (2001), Abstract; Full Text
Presence of DNA fragmentation and lack of neuroprotective effect in DFF45 knockout mice subjected to traumatic brain injury: A.G. Yakovlev, et al.; Mol. Med. 7, 205 (2001), Abstract;

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