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HTS064M ChemiSCREEN™ Human Angiotensin II Receptor, Type I Membrane Preparation

Human AT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P30556 Entrez Gene Number: NM_000685.4, NM_004835.3, NM_009585.3, NM_031850.2, NM_032049.2
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HTS064M
200 units  
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      Overview

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1AT1
      Description
      Catalogue NumberHTS064M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Human Angiotensin II Receptor, Type I Membrane Preparation
      OverviewFull-length human cDNA encoding AT1
      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, although AT1 is primarily responsible for the effects of Ang II on renal function and development, thirst, electrolyte and water balance, and arterial blood pressure. Binding of Ang II to AT1 activates both Gq and Gi (De Gasparo et al., 2000). Chemicon's AT1 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 AT1 interactions and its ligands. The membrane preparations exhibit a Kd of 0.36 nM for [125I]-Sar1-Ile8-Angiotensin II. With 0.3 nM [125I]-Sar1-Ile8-Angiotensin II, 5 µg/well AT1 Membrane Prep yields greater than 50-fold signal-to-background ratio.
      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 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80oC.
      Quality LevelMQ100
      Applications
      ApplicationHuman AT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay and GTPγS binding.
      Biological Information
      Entrez Gene Number
      Entrez Gene SummaryAngiotensin II is a potent vasopressor 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. This gene encodes the type 1 receptor which is thought to mediate the major cardiovascular effects of angiotensin II. This gene may play role in the generation of reperfusion arrhythmias following restoration of blood flow to ischemic or infarcted myocardium. It was previously thought that a related gene, denoted as AGTR1B, existed; however, it is now believed that there is only one type 1 receptor gene in humans. At least four transcript variants have been described for this gene. Additional variants have been described but their full-length nature has not been determined. The entire coding sequence is contained in the terminal exon and is present in all transcript variants.
      Gene Symbol
      • AGTR1
      • AT2R1A
      • AT2R1B
      • AGTR1B
      • AT2R1
      • AT1BR
      • AGTR1A
      • AT1AR
      • AG2S
      • HAT1R
      • AT1
      • AT1B
      GPCR ClassA
      Host CellsChem-1
      Protein TargetAT1
      Target Sub-FamilyAngiotensin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P30556 # Receptor for angiotensin II. Mediates its action by association with G proteins that activate a phosphatidylinositol- calcium second messenger system.
      SIZE: 359 amino acids; 41061 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Liver, lung, adrenal and adrenocortical adenomas.
      PTM: C-terminal Ser or Thr residues may be phosphorylated.
      DISEASE: SwissProt: P30556 # Defects in AGTR1 are a cause of renal tubular dysgenesis (RTD) [MIM:267430]. RTD is an autosomal recessive severe disorder of renal tubular development characterized by persistent fetal anuria and perinatal death, probably due to pulmonary hypoplasia from early-onset oligohydramnios (the Potter phenotype).
      SIMILARITY: SwissProt: P30556 ## Belongs to the G-protein coupled receptor 1 family.
      Incubation ConditionsMembranes 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 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]-Sar1-Ile8-Angiotensin II. (Perkin Elmer#:NEX-248 )
      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 Sar1-Ile8-Angiotensin II at 0.3 nM
      Product Usage Statements
      Quality AssuranceSignal:background and specific binding values obtained in a competition binding assay with varying amounts of ATaub1 membrane prep:
      table tr th align="left" /td th align="center"10 µg/well/td th align="center"5 µg/well /td /tr tr td align="left" Signal:Background/td td align="center" 115.5/td td align="center" 92.8/td /tr tr td align="left" Specific Binding (cpm) /td td align="center"46,458/td td align="center"43,007 /td /tr /table

      SPECIFICATIONS: 1 unit = 5 µg
      Bmax for [125I] Angiotensin II binding: 5.53 pmol/mg protein;
      Kd for [125I] Angiotensin II binding: ~ 0.36 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
      Global Trade Item Number
      Catalogue Number GTIN
      HTS064M 04053252732324