OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Apoptosis signal-regulating kinase 1 (ASK1) is a serine/threonine protein kinase and identified as a MAPKKK which activates the c-Jun N-terminal kinase (JNK) and p38 MAP kinase pathways and is required for TNFα-induced apoptosis. ASK1 suggests possible mechanisms for redox regulation of the apoptosis signal transduction pathway as well as the effects of antioxidants against cytokine- and stress-induced apoptosis.
Family:
Kinase
Sub Family:
STE
Class:
Protein Ser/Thr
Protein Name:
ASK1
Uniprot Number:
Q99683
Protein Aliases:
apoptosis signal regulating kinase 1
Gene Name:
MAP3K5
Gene ID:
4217
Gene Aliases:
MAPKKK5|ASK1|MEKK5
Accession Number:
NM_005923
Organism:
Human
Construct Details:
649-946 recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometric
Measured Response:
Scintillation
Testing Information
Procedure Summary:
ASK1 (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.33 mg/mL myelin basic protein, 10 mM MgAcetate and [gamma-33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting.
Substrate:
0.33 mg/mL MBP
Tracer:
33P
ATP Concentration:
[ATP]= 200µM (Km= 371µM )
Incubation:
40 min at Room temperature
Control Inhibitor:
Staurosporine
Test Sample Requirements:
Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.
Minimum Order Quantity:
2
Turnaround Time
Standard:
6 Business Days
Service Scheduling Note:
Assay is run on a weekly schedule beginning on Friday. Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Adverse / Beneficial Action:
Inhibition of ASK1 may be useful in the treatment of various cardiovascular, central nervous system and peripheral inflammatory conditions.
Additional Information
Brand:
LeadHunter
Testing Location:
KP - France - Celle L'Evescault
RELATED SOLUTIONS
ASK1 Human STE Kinase Enzymatic Radiometric [10 uM ATP] KinaseProfiler LeadHunter Assay - FR
Item: 14-606KP10
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