ChemiSCREEN Membrane Preparation Recombinant Human V1B Vasopressin Receptor

FUNCTION: SwissProt: P47901 # Receptor for arginine vasopressin. The activity of this receptor is mediated by G proteins which activate a phosphatidyl- inositol-calcium second messenger system.
SIZE: 424 amino acids; 46971 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P47901 ## Belongs to the G-protein coupled receptor 1 family.

Radioligand binding assay, and GTPγS binding.

200 units

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 8-fold signal:background with ¹²⁵I-labeled MCH at 0.4 nM.

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

Maintain frozen at -70$deg;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 50 mM 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: [3H] vasopressin (Perkin Elmer # NET800)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

SPECIFICATIONS: 1 unit = 10 μg membrane preparation
Bmax: 2.79 pmol/mg
Kd: 1.03 nM

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