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HTS193M ChemiSCREEN™ Membrane Preparation Recombinant Human S1P5 Lysophospholipid Receptor

Human S1P5 / EDG8 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

ChemiSCREEN™ Membrane Preparation Recombinant Human S1P5 Lysophospholipid Receptor MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-5 Uni Prot Number: Q9H228 Entrez Gene Number: NM_030760.3
HTS193M
200 units  
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Tabela kluczowych gatunków

SpeciesHost CellsProtein Target
Human Chem-5S1P5 / EDG8
Description
Catalogue NumberHTS193M
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human S1P5 Lysophospholipid Receptor
OverviewFull-length human EDG8 cDNA encoding S1P5
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 (Spiegel and Milstien , 2003).
S1P5 can couple with Gi/o and G12/13, and it mediates S1P induced adenylate cyclase inhibition and Ca2+ mobilization like the other S1P receptors. However, unlike the other S1P receptors, it mediates inhibition of MAPK activation and cell proliferation (Im et al., 2000). S1P5 is predominantly expressed in the white matter tracts and oligodendrocytes and is particularly abundant in the anterior commissure, corpus collosum, and optic tract (Terai et al., 2003). S1P induces process retraction in pre-oligodendrocytes and supports cell survival in mature oligodendrocytes by activating S1P5, which indicates a role for S1P5 in maturation and myelination of oligodendrocytes (Jaillard et al., 2005). Millipore's S1P5 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 S1P5 interactions with its ligands. The membrane preparations exhibit EC50s of 2.4 nM for S1P in a GTPγS binding assay.
Product Information
FormatMembranes
PresentationLiquid 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
Quality LevelMQ100
Applications
ApplicationHuman S1P5 / EDG8 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
Entrez Gene SummaryThe lysosphingolipid sphingosine 1-phosphate (S1P) regulates cell proliferation, apoptosis, motility, and neurite retraction. Its actions may be both intracellular as a second messenger and extracellular as a receptor ligand. S1P and the structurally related lysolipid mediator lysophosphatidic acid (LPA) signal cells through a set of G protein-coupled receptors known as EDG receptors. Some EDG receptors (e.g., EDG1; MIM 601974) are S1P receptors; others (e.g., EDG2; MIM 602282) are LPA receptors.[supplied by OMIM]
Gene Symbol
  • EDG8
  • SPPR-2
  • SPPR-1
  • Edg-8
  • S1P5
  • S1PR5
GPCR ClassA
Host CellsChem-5
Protein TargetS1P5 / EDG8
Target Sub-FamilyLysophospholipid
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q9H228 # 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. Is coupled to both the G(i/0)alpha and G(12) subclass of heteromeric G-proteins (By similarity). May play a regulatory role in the transformation of radial glial cells into astrocytes and may affect proliferative activity of these cells.
SIZE: 398 amino acids; 41775 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Widely expressed in the brain, most prominently in the corpus callosum, which is predominantly white matter. Detected in spleen, peripheral blood leukocytes, placenta, lung, aorta, and fetal spleen. Low-level signal detected in many tissue extracts. Isoform 1 is predominently expressed in peripheral tissues, whereas isoform 2 is expressed in brain, spleen and peripheral blood leucocytes.DEVELOPMENTAL STAGE: At 24 weeks of gestation, fragments of radial glial fibers are positive within the cortical plate and subplate of allocortical areas. These positive fragments often appear enlarged as varicosities and some of them terminate at blood vessels. Between 28 and 30 weeks of gestation, all iso- and allocortical areas contain immunolabelled radial glial fibers revealing curvature next to sulci. After 32 weeks of gestation, radial glial fibers gradually disappear; instead positive transitional stages between radial glia and astrocytes were found.DISEASE:SwissProt: Q9H228 # Overexpressed in leukemic large granular lymphocyte (LGL). LGL is a lymphopropliferative disorder often associated with autoimmune disease.
SIMILARITY: SwissProt: Q9H228 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsASSAY CONDITIONS: 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 MgCl2/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled S1P 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.

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 S1P5 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.
Product Usage Statements
Quality Assurance
SPECIFICATIONS: 1 unit = 5 µg membrane preparation

EC50 in GTPγS binding assay by S1P: ~ 2.4 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
Global Trade Item Number
Numer katalogowy GTIN
HTS193M 04053252400780