Caspase 1, recombinant human active

General description

Caspase-1 (also known as ICE, Interleukin-1beta-Converting Enzyme) is the prototypical member of the caspase family of cysteine proteases. Caspase-1 exists in cells as an inactive 45 kDa pro-enzyme. The pro-enzyme is matured by proteolysis to yield large (20 kD) and small (10 kD) subunits. The active enzyme is a heterotetramer consisting of two large and two small subunits. To date the mechanism of regulation of caspase-1 activation is complex and poorly understood. In THP-1 cells, a large proportion of the caspase-1 is present in the inactive pro-enzyme form.

The active recombinant human caspase-1 was expressed in E. coli. The expressed caspase-1 spontaneously undergoes autoprocessing to yield the subunits characteristic of the native enzyme. The recombinant caspase-1 preferentially cleaves the substrates consisting of consensus sequence YVAD (e.g., YVAD-AFC and YVAD-pNA). The recombinant enzyme can also be used as a positive control in caspase assays or in determining the specificity of caspase substrates.

Recommended usage: 0.5-1 unit for fluorometric caspase assays and 2-4 units for colorimetric caspase assays. Optimal working dilutions must be determined by end user.

Physical form

Lyophilized. Reconstitute in 25 μL PBS containing 15% glycerol.

Storage and Stability

Maintain unopened and rehydrated product at -70°C in undiluted, freeze-thaw protected, tightly sealed aliquots. Avoid repeated freeze/thaw cycles. Product stable for 6 months from date of purchased if handled appropriately.

Analysis Note

Partially purified recombinant caspase 3 with a full length HIS-6-tag. Approximately 10-20% caspase 1 protein. The remainder is E.Coli proteins. Approximately 0.8-2 mg total protein/U of activity; E.Coli proteins have no reactivity.

Legal Information

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

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.