ChemiScreen™ Membrane Preparation
Recombinant Human V1B Vasopressin Receptor

ChemiScreen™ Membrane Preparation
Recombinant Human V1B Vasopressin Receptor

• ChemiScreen
• Chemicon (Millipore)

2 mL

Human V1B
HOST CELLS: Chem-1, an adherent mammalian cell line without any endogenous V1B expression.

Arginine vasopressin (AVP) is a 9 amino acid peptide that functions as an antidiuretic, vasoconstrictor and neurotransmitter. The three vasopressin receptors, V1A, V1B and V2, are GPCRs; V1A and V1B couple to Gq and calcium release, whereas V2 couples to Gs (Birnbaumer, 2000). The V1B receptor is expressed prominently in the anterior pituitary, where it mediates vasopressin-induced release of ACTH (Tanoue et al., 2004). A selective antagonist of V1B has recently been developed and shown to reduce depression, anxiety, and aggression in rodents (Blanchard et al, 2005; Griebel et al., 2002). Chemicon's cloned human V1B-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant V1B expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. Chemicon's V1B membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of V1B interactions with vasopressin. The membrane preparations exhibit a Kd of 1.03 nM for [³H]-vasopressin. With 10 ug/well V1b Membrane Prep and 1.5 nM [³H]-vasopressin, a greater than 4-fold signal-to-background ratio was obtained.

Radioligand binding assay, and GTPγS binding.

Human Only

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

The protein encoded by this gene acts as receptor for arginine vasopressin. This receptor belongs to the subfamily of G-protein coupled receptors which includes AVPR1A, V2R and OXT receptors. Its activity is mediated by G proteins which stimulate a phosphatidylinositol-calcium second messenger system. The receptor is primarily located in the anterior pituitary, where it stimulates ACTH release. It is expressed at high levels in ACTH-secreting pituitary adenomas as well as in bronchial carcinoids responsible for the ectopic ACTH syndrome. A spliced antisense transcript of this gene has been reported but its function is not known.

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.

Human

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

Chem-1

Chemicon

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.

Chem-1

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 4-fold signal:background with 3H-labeled vasopressin at 1.5 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&edg;C.

V1B

P47901

peptide

Vasopressin

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

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A

200 units

• AVPR1B
• VPR3
• V1bR
• AVPR3

GPCR Membrane Preparations

ChemiScreen™ Membrane Preparation
Recombinant Human V1B Vasopressin Receptor

NM_000707.2

GPCR