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 inflamatory 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 8 (FLICE, Mch5 and MACH) is an upstream caspase, which can activate all other caspases. Caspase 8 becomes activated through its interaction with the adaptor protein FADD. FADD binds caspase 8 when the cytokine receptor CD95 is activated. Caspase 8 is constitutively expressed as a 55 kD protein and, when activated, generates 18 and 17 Kd mature polypetides. Mice deficient in caspase 8 die in uterus due to impaired heart muscle development and congested accumulation of erythrocytes. Substrates of caspase 8 include caspase 3, 7 and 13. Caspase inhibitors are currently sought as therapeutics for such diseases as Alzheimer's, Parkinson's and cerebral/myocardial ischemia.
Family:
Protease
Sub Family:
Caspase
Class:
Cysteine Peptidase
Protein Name:
Caspase 8
Uniprot Number:
Q14790
Gene Name:
CASP8
Gene ID:
841
Gene Aliases:
MCH5|MACH|FLICE|Casp-8
Accession Number:
NM_001228
Organism:
Human
Construct Details:
Fragment (217-end), 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 8 expressed in E. coli is used. Test compound and/or vehicle is preincubated with 340 ng/ml enzyme‡ in modified HEPES buffer pH 7.4 for 15 minutes at 37⁰C. The reaction is initiated by addition of 50 μM Ac-IEVD-AMC for another 60 minute incubation period. Determination of the amount of AMC formed is read spectrofluorimetrically at 360 nm/465 nm. Compounds are screened at 10 μM.
Substrate:
50 µM Ac-IETD-AMC
Incubation:
60 min at 37°C
Control Inhibitor:
Ac-IETD-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:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
RELATED SOLUTIONS