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Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets.

ed with the appropriate PerkinElmer radioligand, thereby making ScreenReady Targets a complete assay solution for high throughput screening (HTS) campaigns, focused library screening or small to large scale profiling (Figure 1). Twenty-one different ScreenReady Targets are currently available from PerkinElmer, one of which is the human motilin MTL-R1A receptor. Motilin is a 22-amino acid peptide secreted from endocrine cells of the small intestine that stimulates gastrointestinal contractility (Tonini et al ., 1996). The receptor for motilin has been cloned recently (Feighner et al ., 1999) and belongs to the GPCR family. Development of motilin receptor agonists and antagonists would be useful in the treatment of multiple disorders of the gastrointestinal motility. Here we present the pharmacological properties of the human motilin MTL-R1A receptor using the motilin ScreenReady Targets product.

Material & Methods

Material
[125I]-motilin (2200 Ci/mmole) and 96-well FlashPlate Basic microplates were from PerkinElmer Life Sciences Inc.

Motilin was obtained from Bachem Inc. Membranes from CHO cells stably expressing the human motilin MTL-R1A receptor were used for the study.

Methods
Assay Format
Membranes from CHO cells expressing the human motilin MTL-R1A receptor were immobilized into 96-well FlashPlate Basic microplates using PerkinElmers proprietary coating procedure. Stability studies were performed using motilin ScreenReady Targets stored for various times at 4C in the dark. Homogeneous radioligand assays were performed in a volume of 25 μl.

Homogeneous Radioligand Binding Assay
Experiments were performed in binding buffer (50 mM Tris-HCl pH 7.4, 10 mM MgCl2, 1 mM EDTA, 0.1% BSA) using a two-step procedure. Brie
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