HTS208M

ChemiSCREEN Membrane Preparation Recombinant Human AT₂ Angiotensin II Receptor

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

Propiedades

• description: GPCR Class A; protein target AT2
• Quality Level: 100
• technique(s): radioligand binding assay (RLBA): suitable
• UniProt accession no.: P50052
• Gene Information: human ... AGTR2(186)

Descripción

General description

FUNCTION: SwissProt: P50052 # Receptor for angiotensin II. Cooperates with MTUS1 to inhibit ERK2 activation and cell proliferation.
SIZE: 363 amino acids; 41184 Da
SUBUNIT: Interacts with MTUS1.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: In adult, highly expressed in myometrium with lower levels in adrenal gland and fallopian tube. Very highly expressed in fetal kidney and intestine.
SIMILARITY: SwissProt: P50052 ## Belongs to the G-protein coupled receptor 1 family.

Application

Radioligand binding assay

Physical form

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

Storage and Stability

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

Analysis Note

RECOMMENDED ASSAY CONDITIONS: 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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) 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]-CGP42112. (Perkin Elmer # NEX-324 )

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 50-fold signal:background with ¹²⁵I labeled CGP42112 at 0.25 nM

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.