HTS136M

ChemiSCREEN Membrane Preparation Recombinant Human V1B Vasopressin Receptor

• UNSPSC Code: 41123201
• eCl@ss: 32161000
• NACRES: NA.45

Properties

• description: GPCR Class A; protein target V1B
• Quality Level: 100
• technique(s): radioligand binding assay (RLBA): suitable
• UniProt accession no.: P47901
• Gene Information: human ... AVPR1B(553)

Description

General description

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.

Application

Radioligand binding assay, and GTPγS binding.

Packaging

200 units

Quality

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

Physical form

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.

Storage and Stability

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

Analysis Note

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.

Disclaimer

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.