merck millipore,默克密理博,HTS068M,ChemiSCREEN™ Membrane Preparation Recombinant Human APJ Apelin Receptor

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Merck Millipore 默克密理愽

应用说明	品种	宿主细胞	蛋白靶
Radioligand binding assay and GTPγS binding.	Human	Chem-1	APJ

描述
产品目录编号	HTS068M
商名	Chemicon ChemiScreen
描述	ChemiSCREEN™ Membrane Preparation Recombinant Human APJ Apelin Receptor
概述	Full-length human AGTRL1 cDNA encoding APJ
背景信息	Apelin peptides have been discovered to be a family of peptides of different sizes that is derived from the N-terminus of a 77 amino acid precursor peptide (preproapelin) (Hosoya et al., 2000). Apelin receptor (APJ) is a G protein-coupled receptor that is activated by several apelin fragments. APJ activation leads to the inhibition of cAMP production. APJ and apelin peptides have been found to be involved in the regulation of cardiovascular function (Katugampola et al., 2001) and fluid homeostasis (Reaux et al., 2001). Broad roles of apelin system has been established in lowering blood pressure, as a potent cardiac inotrope, in modulating pituitary hormone release and food and water intake, in stress activation, and as a novel adipokine that is excreted from fat cells and regulates insulin (Lee et al., 2006) . Millipore's APJ 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 APJ. The membrane preparations exhibit a Kd of 0.4 nM for [¹²⁵I]-Apelin. With 0.25 nM [¹²⁵I]-Apelin, 5 g/well APJ Membrane Prep typically yields greater than 20-fold signal-to-background ratio.

产品信息
格式	Membranes
演示	Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives. Packaging method: Membranes protein were adjusted to the indicated concentration in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.

应用
应用	Human APJ GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
应用说明	Radioligand binding assay and GTPγS binding.

生物信息
品种	Human
Entrez基因编号	NP_005152
GPCR类别	A
宿主细胞	Chem-1
蛋白靶	APJ
靶子系列	Apelin
UniProt编号	P35414
孵化条件	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]-Apelin. (Perkin Elmer#:NEX-393 ) 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 20-fold signal:background with ¹²⁵I labeled Apelin at 0.25 nM

产品使用声明

质量保证

Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of APJ membrane prep.

	10 µg/well	5 µg/well
Signal:Background	33	50
Specific Binding (cpm)	17297	15015

SPECIFICATIONS: 1 unit = 5 µg membrane preparation
B_max for [¹²⁵I]-Apelin binding: 3.85 pmol/mg protein
K_d for [¹²⁵I]-Apelin binding: ~0.4 nM

使用声明

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.

储存和货运信息
存储条件	Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

包装信息
数量	200 units

merck millipore,默克密理博,HTS068M,ChemiSCREEN™ Membrane Preparation Recombinant Human APJ Apelin Receptor

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