ASK1 Human STE Kinase Enzymatic Radiometric LeadHunter Assay [10 μM ATP] - TW

Item: 167150

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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:

Fragment (649-946), Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Radiometry

Measured Response:

Scintillation

Testing Information

Procedure Summary:

Human recombinant protein kinase ASK1 expressed in E. coli is used. Test compound and/or vehicle is preincubated with 0.14 U/ml enzyme‡ in modified MOPS buffer pH 7.2 for 15 minutes at 37°C. The reaction is initiated by addition of 50 µg/ml myelin basic protein (MBP), 10 µM ATP and 0.25 µCi [γ32P]ATP for another 30 minute incubation period and terminated by further addition of 3% H3PO4. An aliquot is removed and counted to determine the amount of [32P]MBP formed. Compounds are screened at 10 µM.

Substrate:

50 µg/ml Myelin basic protein (MBP)

ATP Concentration:

10 uM

Incubation:

30 min at 37°C

Control Inhibitor:

Staurosporine

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:

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

122130844

Name

Staurosporine

Measurement

0.037μM - IC50

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:

Taiwan - Taipei