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merck millipore,默克密理博,HTS010M,ChemiSCREEN™ Membrane Preparation Recombinant Rehsus Macaque CCR5 Chemokine Receptor
产品名称：ChemiSCREEN™ Membrane Preparation Recombinant Rehsus Macaque CCR5 Chemokine Receptor
产品型号：HTS010M
Rhesus Macaque CCR5 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
merck millipore,默克密理博,HTS010M,ChemiSCREEN™ Membrane Preparation Recombinant Rehsus Macaque CCR5 Chemokine Receptor
- 产品介绍
- merck millipore,默克密理博,HTS010M,ChemiSCREEN™ Membrane Preparation Recombinant Rehsus Macaque CCR5 Chemokine Receptor
重要规格表
应用说明 品种 宿主细胞 蛋白靶 Radioligand binding assay and GTPγS binding. Rhesus Macaque Chem-1 CCR5 描述 产品目录编号 HTS010M 商名 - Chemicon
- ChemiScreen
描述 ChemiSCREEN™ Membrane Preparation Recombinant Rehsus Macaque CCR5 Chemokine Receptor 概述 Full-length rhesus macaque CCR5 cDNA encoding CCR5 背景信息 CCR5 is the receptor for CC chemokines MIP-1α, MIP-1β, and RANTES (Raport et al., 1996), and is preferentially expressed on Th1 lymphocytes (Loetscher et al., 1998). CCR5 is a coreceptor for macrophage-tropic HIV, and its ligands potently inhibit HIV replication in human leukocytes (Cocchi et al., 1995). In addition, HIV-infected patients with the nonfunctional CCR5Δ32 allele exhibit delayed onset of AIDS symptoms (Samson et al., 1996), and pharmacological antagonism of CCR5 inhibits HIV-1 infection (Strizki et al., 2001). Preclinical testing of small molecule antagonists of CCR5 has been hampered by low affinity of the compounds to rodent and dog CCR5, but two such compounds, maraviroc and AD101, have been shown to have potent antagonist activity at rhesus macaque CCR5, which differs from human CCR5 by 8 amino acids (Napier et al., 2005; Billick et al., 2004). One antagonist of human CCR5, SCH-C, does not block HIV entry through rhesus macaque CCR5, and one amino acid difference is responsible for the functional difference (Billick et al., 2004). Millipore's rhesus macaque CCR5 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 rCCR5. The membrane preparations exhibit a Kd of 0.32 nM for [125I]-MIP-1α. With 0.25 nM [125I]-MIP-1α, 5 μg/well rCCR5 Membrane Prep typically yields greater than 7-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 no preservatives.
Packaging method: Membranes protein were adjusted to 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C应用 应用 Rhesus Macaque CCR5 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 Rhesus Macaque Entrez基因编号 - NM_001042773.2
GPCR类别 A 宿主细胞 Chem-1 蛋白靶 CCR5 靶子系列 Chemokine 孵化条件 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]-MIP-1α (Perkin Elmer#: NEX-298 )
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 7-fold signal:background with 125I labeled MIP-1α at 0.15 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 CCR5 Receptor membrane prep.
10 µg/well 5 µg/well Signal:Background 13.6 12.1 Specific Binding (cpm) 5869 4538
SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax for [125I]-MIP-1α binding: 1.9 pmol/mg protein
Kd for [125I]-MIP-1α binding: ~0.35 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
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