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Performance of the AMARA HitHunter Chemiluminescence Kinase Assay with the LEADseeker Multimodality Imaging System

Key words: AMARA • kinase assay • LEADseeker • chemiluminescence


It is predicted that at least 500 kinase and phosphatase enzymes exist, their activities having wide-ranging regulatory responses. Many kinases are part of a signaling cascade in disease pathways such as inflammation and cancer, so identifying modulators of kinase and phosphatase activity may provide new therapeutic agents.

This application note provides recommendations for the successful imaging of the AMARA HitHunter™ Chemiluminescence Kinase Assay on the LEADseeker™ Multimodality Imaging System and demonstrates the assay’s ability to measure the functional activity of serine/threonine kinase AMP in the phosphorylation of the AMARA substrate (AMARAASAAALARRR).


Materials
Reagents and instrumentation

AMARA HitHunter Chemiluminescence 90-0060-02
Kinase Assay Kit*, 800 (384 wells)

LEADseeker Multimodality Imaging System 18-1140-71

[14C] Uniform Reference Plate CFQ11387-4UCI


Additional materials required
AMP-activated protein kinase (Upstate #14-305)

ATP (Sigma A-7699)

Adenosine monophosphate (AMP) (Sigma A-2002)

384-well white flat bottom polystyrene (Corning 3705)
not treated microplates

Graphpad Prism software v.4 (GraphPad
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