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HTS176M ChemiSCREEN™ Membrane Preparation Recombinant Human S1P1 Lysophospholipid Receptor

Human S1P1 / EDG1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P21453 Entrez Gene Number: NM_001400
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HTS176M
200 units  
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Overview

Replacement Information

Key Specifications Table

SpeciesHost CellsProtein Target
Human Chem-1S1P1 / EDG1
Description
Catalogue NumberHTS176M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human S1P1 Lysophospholipid Receptor
OverviewFull-length human EDG1 cDNA encoding S1P1
Background InformationSphingosine 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.
References
Product Information
FormatMembranes
PresentationLiquid 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.
Quality LevelMQ100
Applications
ApplicationHuman S1P1 / EDG1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay and GTPγS binding
Biological Information
Entrez Gene Number
Gene Symbol
  • S1PR1
  • CHEDG1
  • EDG1
GPCR ClassA
Host CellsChem-1
Protein TargetS1P1 / EDG1
Target Sub-FamilyLysophospholipid
UniProt Number
Incubation ConditionsMembranes 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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance1 unit = 5 μg
EC50 in GTPγS binding assay by Sphingosine 1-phosphate: ~ 9.5 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 ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
HTS176M 04053252581120

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