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HTS097M Sigma-AldrichChemiSCREEN™ Membrane Preparation Recombinant Human S1P₃ Lysophospholipid Receptor

Human S1P3 / EDG3 GPCR membrane preparation for GTPγS Binding & Radioligand Binding Assays.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

MSDS
参考

Host Cell: Chem-1 Uni Prot Number: Q99500 Entrez Gene Number: NM_005226.2

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概述

Replacement Information

重要规格表

Species	Host Cells	Protein Target
Human	Chem-1	S1P3 / EDG3

Description
Catalogue Number	HTS097M
Trade Name	ChemiScreen Chemicon
Description	ChemiSCREEN™ Membrane Preparation Recombinant Human S1P₃ Lysophospholipid Receptor
Overview	Full-length human EDG3 cDNA encoding S1P₃
Background Information	Sphingosine 1-phosphate (S1P) is a bioactive lipid that binds to and activates a family of GPCRs, S1P_1-5 (also known as EDG receptors). Interactions between S1P and its receptors mediate cytoskeletal rearrangement and cell migration, with functional consequences in angiogenesis, lymphocyte trafficking, and smooth muscle development (Anliker and Chun, 2004). S1P₁ (Edg-1) signals exclusively through G_i, whereas S1P₂ (Edg-5) and S1P₃ (Edg-3) activate G_i, G_q and G_12/13 (Windh et al., 1999). Although S1P₁ and S1P₃ promote cell migration, S1P₂ inhibits cell migration in several cell types; these opposing functions appear to result from differences in the ability of each receptor to activate G_i (Sugimoto et al., 2003). Studies with knockout mice indicate that S1P₂ and S1P₃ have redundant functions in maintaining vascular integrity during embryonic development (Kono et al., 2004). In addition, S1P₃ regulates immune responses by contributing to endothelial barrier function in splenic marginal zones (Girkontaite et al., 2004). Millipore's S1P₃ 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 S1P₃ interactions with its ligands. The membrane preparation exhibits EC50 of 1.7nM for S1P in a GTPγS binding assay.

References

Product Information
Format	Membranes
Presentation	Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol with no preservatives. Packaging method: Membrane protein was adjusted to 1 mg/ml in packaging buffer, rapidly frozen, and stored at -80°C
Quality Level	MQ100

Applications
Application	Human S1P3 / EDG3 GPCR membrane preparation for GTPγS Binding & Radioligand Binding Assays.
Key Applications	GTPγS binding Radioligand binding assay
Application Notes	GTPγS Binding and Radioligand Binding Assay.

Biological Information
Entrez Gene Number	NM_005226.2
Entrez Gene Summary	This gene encodes a member of the EDG family of receptors, which are G protein-coupled receptors. This protein has been identified as a functional receptor for sphingosine 1-phosphate and likely contributes to the regulation of angiogenesis and vascular endothelial cell function.
Gene Symbol	EDG3 S1P3 LPB3 FLJ37523 EDG-3 S1PR3 MGC71696
GPCR Class	A
Host Cells	Chem-1
Protein Target	S1P3 / EDG3
Target Sub-Family	Lysophospholipid
UniProt Number	Q99500
UniProt Summary	FUNCTION: SwissProt: Q99500 # Receptor for the lysosphingolipid sphingosine 1- phosphate (S1P). S1P is a bioactive lysophospholipid that elicits diverse physiological effect on most types of cells and tissues. When expressed in rat HTC4 hepatoma cells, is capable of mediating S1P-induced cell proliferation and suppression of apoptosis. SIZE: 378 amino acids; 42250 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. TISSUE SPECIFICITY: Expressed in all tissues, but most abundantly in heart, placenta, kidney, and liver. SIMILARITY: SwissProt: Q99500 ## Belongs to the G-protein coupled receptor 1 family.
Incubation Conditions	ASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [³⁵S]-GTPγS (final concentration of 0.1 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl₂/0.5 µM GDP in a nonbinding 96-well plate. Unlabeled S1P is 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, and washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4. The plate is dried and counted. 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 S1P -stimulated [³⁵S]-GTPγS binding. The S1P₃ 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.

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	EC50 in GTPγS binding assay by S1P: ~ 1.7nM
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.

Packaging Information
Material Size	200 units

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
产品目录编号	GTIN
HTS097M	04053252405297

Documentation

ChemiSCREEN™ Membrane Preparation Recombinant Human S1P₃ Lysophospholipid Receptor MSDS

职位
物料安全数据表 (MSDS)

参考

参考概述	公共医疗ID
Lysophospholipid G protein-coupled receptors. Anliker, Brigitte and Chun, Jerold J. Biol. Chem., 279: 20555-8 (2004) 2004 显示摘要 The many biological responses documented for lysophospholipids that include lysophosphatidic acid and sphingosine 1-phosphate can be mechanistically attributed to signaling through specific G protein-coupled receptors. At least nine receptors have now been identified, and the total number is likely to be larger. In this brief review, we note cogent features of lysophospholipid receptors, including the current nomenclature, signaling properties, development of agonists and antagonists, and physiological functions.	15023998
The Sphingosine-1-phosphate Receptors S1P1, S1P2, and S1P3 Function Coordinately during Embryonic Angiogenesis. Kono M et al. J. Biol. Chem., 279:29367-29373 (2004) 2004	15138255
The sphingosine-1-phosphate (S1P) lysophospholipid receptor S1P3 regulates MAdCAM-1+ endothelial cells in splenic marginal sinus organization Girkontaite, Irute, et al J Exp Med, 200:1491-501 (2004) 2004	15583019
Inhibitory and Stimulatory Regulation of Rac and Cell Motility by the G12/13-Rho and Gi Pathways Integrated Downstream of a Single G Protein-Coupled Sphingosine-1-Phosphate Receptor Isoform. Sugimoto N et al. Mol. Cell. Biol., 23:1534-1545 (2003) 2003	12588974
Differential coupling of the sphingosine 1-phosphate receptors Edg-1, Edg-3, and H218/Edg-5 to the G(i), G(q), and G(12) families of heterotrimeric G proteins Windh, R T, et al J Biol Chem, 274:27351-8 (1999) 1999	10488065

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