Biological Information
Background Information:
Caspases 9 is a member of the CED-3/caspase-3 subfamily. Upon activation, it functions to activate downstream effector caspases in the apoptotic pathway. 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 9
Uniprot Number:
P55211
Protein Aliases:
protein phosphatase 1, regulatory subunit 56
Gene Name:
CASP9
Gene ID:
842
Gene Aliases:
MCH6|ICE-LAP6|APAF-3|PPP1R56
Accession Number:
NM_032996
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 9 expressed in E. coli is used. Test compound and/or vehicle is preincubated with 10.2 µg/mL enzyme in modified HEPES buffer pH 7.4 for 15 minutes at 37°C. The reaction is initiated by addition of 25 µM Ac-LEHD-AFC for another 60 minutes incubation period. Determination of the amount of AFC formed is read spectrofluorimetrically at 400 nm/510 nm. Compounds are screened at 10 µM.
Substrate:
25 µM Ac-LEHD-AFC
Incubation:
60 min at 37°C
Control Inhibitor:
Ac-LEVD-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
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