ChemiScreen™ Membrane Preparation Recombinant Hu Angiotensin II Receptor, Type 1

Species	Host Cells	Protein Target
Human	Chem-1	AT1

Species

Host Cells

Protein Target

Human

Chem-1

AT1

Description
Catalogue Number	HTS064M
Brand Family	Chemicon®
Trade Name	ChemiScreen Chemicon
Description	ChemiSCREEN™ Human Angiotensin II Receptor, Type I Membrane Preparation
Overview	Full-length human cDNA encoding AT₁
Background Information	Angiotensin 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 G_q and G_i (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.

Description

Catalogue Number

HTS064M

Brand Family

Chemicon®

Trade Name

ChemiScreen
Chemicon

Description

ChemiSCREEN™ Human Angiotensin II Receptor, Type I Membrane Preparation

Overview

Full-length human cDNA encoding AT₁

Background Information

Angiotensin 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 G_q and G_i (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
Format	Membranes
Presentation	Liquid 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.

Product Information

Format

Membranes

Presentation

Liquid 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.

Applications
Application	Human AT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications	Radioligand binding assay GTPγS binding
Application Notes	Radioligand binding assay and GTPγS binding.

Applications

Application

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

Key Applications

Radioligand binding assay
GTPγS binding

Application Notes

Radioligand binding assay and GTPγS binding.

Biological Information
Entrez Gene Number	NM_000685.4 NM_004835.3 NM_009585.3 NM_031850.2 NM_032049.2
Entrez Gene Summary	Angiotensin 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 Class	A
Host Cells	Chem-1
Protein Target	AT1
Target Sub-Family	Angiotensin
UniProt Number	P30556
UniProt Summary	FUNCTION: 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 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, 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

Biological Information

Entrez Gene Number

Entrez Gene Summary

Angiotensin 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 Class

Host Cells

Chem-1

Protein Target

AT1

Target Sub-Family

Angiotensin

UniProt Number

P30556

UniProt Summary

FUNCTION: 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 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, 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 Assurance	Signal:background and specific binding values obtained in a competition binding assay with varying amounts of AT aub 1 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.

Product Usage Statements

Quality Assurance

Signal:background and specific binding values obtained in a competition binding assay with varying amounts of AT aub 1 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 Conditions	Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Storage and Shipping Information

Storage Conditions

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

Packaging Information
Material Size	200 units

Packaging Information

Material Size

200 units

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

办公用品

ChemiScreen™ Membrane Preparation Recombinant Hu Angiotensin II Receptor, Type 1

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