ChemiSCREEN Recombinant Human PRP/GPR10 Prolactin-Releasing Peptide Receptor Membrane Preparation

FUNCTION: SwissProt: P49683 # Receptor for prolactin-releasing peptide (PrRP). Implicated in lactation, regulation of food intake and pain-signal processing.
SIZE: 370 amino acids; 41107 Da
SUBUNIT: Interacts through its C-terminal region with the PDZ domain-containing proteins GRIP1, GRIP2 and PICK1. Interacts with PDZ domains 4 and 5 of GRIP1 and with the PDZ domain of PICK1.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Only detected in the pituitary gland and in all cell types of pituitary adenomas.
SIMILARITY: SwissProt: P49683 ## Belongs to the G-protein coupled receptor 1 family.

Radioligand binding assay, and GTPγS binding.

1 mL, 1 mg/mL

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 30-fold signal:background with ¹²⁵I-labeled PRP-20 at 0.25 nM.

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl₂, 1 mM CaCl₂, 0.2% BSA, filtered and stored at 4°C
Radioligand: [¹²⁵I] PRP-20 (Perkin Elmer # NEX384)
Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.

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