merck millipore,默克密理博,HTS176M,ChemiSCREEN™ Membrane Preparation Recombinant Human S1P1 Lysophospholipid Receptor

搜索"Merck Millipore 密理愽"产品

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	S1P1 / EDG1

描述
产品目录编号	HTS176M
品牌系列	Chemicon®
商名	Chemicon ChemiScreen
描述	ChemiSCREEN™ Membrane Preparation Recombinant Human S1P₁ Lysophospholipid Receptor
概述	Full-length human EDG1 cDNA encoding S1P₁
背景信息	Sphingosine 1-phosphate (S1P) is a biologically active lysophospholipid that transmits signals through a family of five G-protein-coupled receptors to regulate cell proliferation, migration, cytoskeletal organization, and differentiation. S1P₁ was the first identified S1P receptor. It primarily couples to PTX-sensitive Gi/o proteins and mediates S1P-induced adenylate cyclase inhibition. Expression of S1P₁ is pervasive, including spleen, brain, heart, lung, adipose tissues, liver, thymus, kidney, and skeletal muscle (Zhang et al. 1999). The deletion of S1P₁ in mice results in embryonic lethality (Liu et al., 2000) with death attributable to incomplete vascular maturation. Recent reports demonstrate specific roles for S1P₁ in lymphocyte recirculation/egress (Matloubian et al., 2004). Millipore's S1P₁ 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 S1P₁ interaction with its ligands. The cell line exhibits a calcium response with EC50s of 141 μM for Sphingosine 1-phosphate. The membrane preparations exhibit EC50s of 9.5 μM for Sphingosine 1-phosphate in a GTPγS binding assay.

产品信息
演示	Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives. Packaging method: Membrane protein was adjusted to 1 mg/ml in packaging buffer, rapidly frozen, and stored at -80°C.

应用
应用	Human S1P1 / EDG1 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	NM_001400
GPCR类别	A
宿主细胞	Chem-1
蛋白靶	S1P1 / EDG1
靶子系列	Lysophospholipid
孵化条件	Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [³⁵S]-GTPγS (final concentration of 0.3 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl₂/10 μM GDP in a nonbinding 96-well plate. Unlabeled Sphingosine 1-phosphate was added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30°C. The binding reaction is transferred to a GF/B filter plate (Millipore MAHF B1H) previously prewetted with water. The plate is washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4, then dried and counted. Note: Performing the reaction with lower GDP concentrations (0.5 μM) results in elevated constitutive activity (Figure 2). One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific sodium propionate-stimulated [³⁵S]-GTPγS binding. The S1P1 membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.

产品使用声明
质量保证	1 unit = 5 μg EC50 in GTPγS binding assay by Sphingosine 1-phosphate: ~ 9.5 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,HTS176M,ChemiSCREEN™ Membrane Preparation Recombinant Human S1P1 Lysophospholipid Receptor

whatman,沃特曼,nalgene,耐洁,merck millipore,默克密理愽,sartorius,赛多利斯,pall,颇尔
thermo fisher,赛默飞世尔,GE,MN,macherey-nagel,advantec,wheaton,brand,普兰德,welch,airtech