AMPK Human PRKAA1 Kinase Enzymatic Radiometric Activation LeadHunter Assay [1 μM ATP] - TW

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OVERVIEW

DOCUMENTS/REFERENCES

Biological Information

Background Information:

Adenosine 5’-monophosphate (AMP)-activated protein kinase (AMPK) is a metabolic master switch, activated by exercise and muscle contraction. AMPK phosphorylate key target proteins that control glucose uptake, oxidation of fatty acids, cholesterol synthesis, lipogenesis, triglyceride synthesis, adipocyte lipolysis, glycolysis and nitric oxide synthesis. Metabolic deficiencies or inhibition of AMPK signaling system may result in impaired glucose tolerance and Type 2 diabetes mellitus and contribute to lipid accumulation.

Family:

Kinase

Protein Name:

AMPK

Uniprot Number:

Q13131　

Protein Aliases:

Acetyl-CoA carboxylase kinase　

Accession Number:

NM_006251

Organism:

Human

Construct Details:

Full Length, Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Agonist

Detection Method:

Radiometry

Measured Response:

Radiometry

Testing Information

Procedure Summary:

Human recombinant protein kinase AMPK (A1/B1/G1) expressed in insect cells is used. Test compound or 1 µM A769662 or vehicle is preincubated with 0.24 µg/ml of enzyme in modified MOPS buffer pH 7.2 for 15 minutes at 37C. The reaction is initiated by addition of 10 mM SAMS peptide, 1 mM ATP and 0.25 mCi [g32P]ATP for another 30 minute incubation period and terminated by further addition of 3% H3PO4. After harvesting, filtermats are counted to determine the amount of [32P]SAMS peptide formed. Compounds are screened at 10 µM.

Substrate:

10 µM SAMS peptide

Incubation:

30 min at 37°C

Control Inhibitor:

A769662

Criteria For Significance:

≥ 50% of max stimulation

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

Clinical Relevance

Therapeutic Area:

Metabolic Diseases

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei