Analysis Note

Also contains E. coli proteins.Approximately, 0.8-2 µg total protein / unit of activity.

Application

Enzyme is prepared as an activity standard only, any other application not tested. Caspase-9 is subject to auto-activation thus active caspase subunits and partially active intermediates will be seen. Bands of approximately 45kDa (pro), 35kDa (partially active) and 18kDa + 12kDa (large and small subunits) may be seen by western blot depending upon the antibody used. The small subunit is extremely difficult to detect in most westerns.

Activation: incubate 1-5 µL of stock in PBS pH 7.5 (100mM HEPES can be used as well) plus 10mM ATP and 5mM DTT (added just before use) for 30 minutes at 37°C.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Product Source: Human

Caspase-9 is a member of the interleukin-1beta converting enzyme (ICE) family of cysteine proteases. Similar to other caspases, caspase-9 also exists in cells as an inactive proenzyme. During the initiation of apoptosis the proenzyme is processed at aspartate residues to form active caspase-9. As one of the initiator caspases, active caspase-9 functions to trigger the activation of downstream effector caspases and further lead to disassembly of cell structures.

The partially purified active recombinant human caspase-9 with full-length HIS-6 tag was expressed in E. coli. The expressed caspase-9 spontaneously undergoes autoprocessing to yield the subunits characteristic of the native enzyme. In combination with caspase-9 activity assays, the active recombinant caspase-9 is useful in biological screening of caspase inhibitors. The recombinant enzyme can also be used as a positive control in caspase-9 assays. Caspase 9 is known to have strong reactivity with other sequence peptides, especially DEVD-pNA.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Physical form

Lyophilized. Reconstitute with 25 µl of PBS + 15% glycerol.

Storage and Stability

Maintain at -70°C in undiluted aliquots for 3-6 months from date of receipt. Avoid repeated freeze/thaw cycles.

Unit Definition

Specific Activity: One unit of the recombinant caspase-9 is the enzyme activity that cleaves 1 nmol of the caspase substrate LEHD-pNA (pNA: pnitroanaline) per hour at 37°C at saturated substrate concentrations.