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HTS208M ChemiSCREEN™ Membrane Preparation Recombinant Human AT2 Angiotensin II Receptor

Human AT2 GPCR membrane preparation for Radioligand binding Assays.

Host Cell: Chem-1 Uni Prot Number: P50052 Entrez Gene Number: NM_000686.4
HTS208M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Specifications Table

SpeciesHost CellsProtein Target
Human Chem-1AT2
Description
Catalogue NumberHTS208M
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human AT2 Angiotensin II Receptor
OverviewFull-length human AGTR2 cDNA encoding AT2
Background InformationAngiotensin II (Ang II), an octapeptide produced by cleavage of angiotensinogen by angiotensin-converting enzyme, plays a fundamental role in cardiovascular homeostasis. Two GPCRs, AT1 and AT2, mediate the effects of AngII. Most of the known actions of angiotensin II are mediated through the AT1 receptor and serve to maintain blood pressure and glomerular filtration rate in the face of extracellular volume depletion (Griendling et al. 1996). In vitro and in vivo studies indicated that the AT2 receptor counterbalances the effect of the AT1 receptor (Horiuchi et al. 1999). Expression of the AT2 receptor is developmentally regulated: it is highly expressed in various fetal tissues and at a lower density in adult adrenal medulla, brain, and reproductive tissues (Griendling et al. 1996). AT2 expression appears to be re-expressed or up-regulated after vascular injury, myocardial infarction, cardiac failure or wound healing (Matsubara 1998). Millipore's AT2 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 agonists and antagonists of AT2. The membrane preparations exhibit a Kd of 0.32 nM for [125I]-CGP42112. With 0.25 nM [125I]-CGP42112, 5μg/well AT2 Membrane Prep typically yields greater than 50-fold signal-to-background ratio.
References
Product Information
FormatMembranes
PresentationLiquid 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
Quality LevelMQ100
Applications
ApplicationHuman AT2 GPCR membrane preparation for Radioligand binding Assays.
Key Applications
  • Radioligand binding assay
Application NotesRadioligand binding assay
Biological Information
Entrez Gene Number
Entrez Gene SummaryAngiotensin II is a potent pressor hormone and a primary regulator of aldosterone secretion. It is an important effector controlling blood pressure and volume in the cardiovascular system. It acts through at least two types of receptors termed AT1 and AT2. AGTR2 belongs to a family 1 of G-protein coupled receptors. It is an intergral membrane protein. It plays a role in the central nervous system and cardiovascular functions that are mediated by the renin-angiotensin system. This receptor mediates programmed cell death (apoptosis). In adults, it is highly expressed in myometrium with lower levels in adrenal gland and fallopian tube. It is highly expressed in fetal kidney and intestine. The human AGTR2 gene is composed of three exons and spans at least 5 kb. Exons 1 and 2 encode for 5' untranslated mRNA sequence and exon 3 harbors the entire uninterrupted open reading frame.
Gene Symbol
  • AGTR2
  • MRX88
  • ATGR2
  • AT2
GPCR ClassA
Host CellsChem-1
Protein TargetAT2
Target Sub-FamilyAngiotensin
UniProt Number
UniProt SummaryFUNCTION: 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.
Incubation ConditionsRECOMMENDED 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
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. Signal:background and specific binding values obtained in a competition binding assay with AT2 Receptor membrane prep.
Product Usage Statements
5 µg/well
Signal:Background 148.1
Specific Binding (cpm) 17301

SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax for [125I]-CGP42112 binding: 3.9 pmol/mg protein
Kd for [125I]-CGP42112 binding: ~0.32 nM Usage Statement
  • 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.
Storage and Shipping Information
Storage ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
HTS208M 04053252469534

Documentation

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