Biological Information
Background Information:
Caspases are the executioners of programmed cell death (apoptosis), a normal process used physiologically to eliminate specific cells or cell types without damaging surrounding cells or causing an inflammatory response. All caspases belong to a family of proteases characterized by a cysteine residue in the active site and display specificity for Asp residues. Caspases are present in all cells as a procaspase, which becomes activated through proteolytic cleavage by other proteases including other caspases. Caspase-1 (interleukin-1 beta-converting enzyme, ICE) is responsible for proteolytic activation of the biologically inactive interleukin-1 beta precursor to the proinflammatory cytokine. Inhibition of ICE is a new anti-inflammatory strategy being explored by the design of both reversible inhibitors and irreversible inactivators of the enzyme. ICE inhibitors that cross react against multiple ICE homologs can also block diverse types of apoptosis.
Family:
Protease
Sub Family:
Caspase
Class:
Cysteine Peptidase
Protein Name:
Caspase 1
Uniprot Number:
P29466
Protein Aliases:
caspase-1|interleukin 1, beta, convertase
Gene Name:
CASP1
Gene ID:
834
Gene Aliases:
ICE
Accession Number:
NM_033292
Organism:
Human
Construct Details:
Fragment, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant Caspase 1 expressed in E. coli cells is used. Test compound and/or vehicle are preincubated with 2.5 U/ml‡ of enzyme for 15 minutes at 30⁰C in Hepes buffer pH 7.4. The reaction is initiated by addition of 7.5 µM Z-YUAD-AFC for another 45 minute incubation period. Read plate on a spectrophotometer (Excitation: 405 nm, Emission: 535 nm). Compounds are screened at 10 μM.
Substrate:
7.5 µM Z-YUAD-AFC
Incubation:
45 min at 30°C
Control Inhibitor:
Ac-YVAD-CHO
Criteria For Significance:
≥ 50 of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Inflammation/Allergy
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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