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The development of a melanocortin MC4 receptor binding assay using the LEADseeker Multimodality Imaging System

Key words: Melanocortin • receptor binding assay • LEADseeker • SPA Imaging Beads


Pro-melanocortins are present in high levels in the pituitary and are processed into three major peptide hormone fragments: adrenocorticotrophin (ACTH), the melanocyte stimulating hormones, and β-endorphin. The various melanocyte stimulating hormones are known to activate the MC4 (melanocortin subtype 4) receptor. The MC4 receptor has been linked to the regulation of appetite (1). Evidence has suggested that melanocortins are involved in mediating appetite regulation through the effects of leptin and controlling the expression of neuropeptide Y. Thus, the MC4 receptor is a potential target for anti-obesity agents (2).

This application note describes a 384-well melanocortin MC4 receptor binding assay, which has been developed on the LEADseeker™ Multimodality Imaging System.


Materials
Products used

LEADseeker Multimodality Imaging System 18-1140-71

Wheat Germ Agglutinin (WGA) PEI RPNQ0289
Type B PS SPA Imaging Beads

[125I] (Nle4-D-phe7) α-Melanocyte Stimulating IM316
Hormone ([125I] α-MSH), direct labeled


Other materials required
Human recombinant melanocortin (Euroscreen, ES-191-M)
MC4 receptor membrane preparation*

NDP-α-MSH (Sigma, M-8764)

α-Melanocyte Stimulating Hormone (&alp
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Source:


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