ChemiScreen™ Membrane Prep Hu Recombinant S1P5

Species	Host Cells	Protein Target
Human	Chem-5	S1P5 / EDG8

Species

Host Cells

Protein Target

Human

Chem-5

S1P5 / EDG8

Description
Catalogue Number	HTS193M
Trade Name	ChemiScreen Chemicon
Description	ChemiSCREEN™ Membrane Preparation Recombinant Human S1P₅ Lysophospholipid Receptor
Overview	Full-length human EDG8 cDNA encoding S1P₅
Background Information	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 (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.

Description

Catalogue Number

HTS193M

Trade Name

ChemiScreen
Chemicon

Description

ChemiSCREEN™ Membrane Preparation Recombinant Human S1P₅ Lysophospholipid Receptor

Overview

Full-length human EDG8 cDNA encoding S1P₅

Background Information

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 (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
Format	Membranes
Presentation	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

Product Information

Format

Membranes

Presentation

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

Applications
Application	Human S1P5 / EDG8 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications	Radioligand binding assay GTPγS binding
Application Notes	Radioligand binding assay and GTPγS binding.

Applications

Application

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

Key Applications

Radioligand binding assay
GTPγS binding

Application Notes

Radioligand binding assay and GTPγS binding.

Biological Information
Entrez Gene Number	NM_030760.3
Entrez Gene Summary	The 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 Class	A
Host Cells	Chem-5
Protein Target	S1P5 / EDG8
Target Sub-Family	Lysophospholipid
UniProt Number	Q9H228
UniProt Summary	FUNCTION: 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 Conditions	ASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [³⁵S]-GTPγS (final concentration of 0.3 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.

Biological Information

Entrez Gene Number

NM_030760.3

Entrez Gene Summary

The 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 Class

Host Cells

Chem-5

Protein Target

S1P5 / EDG8

Target Sub-Family

Lysophospholipid

UniProt Number

Q9H228

UniProt Summary

FUNCTION: 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 Conditions

ASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [³⁵S]-GTPγS (final concentration of 0.3 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.

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 Conditions	Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

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

Packaging Information

Material Size

200 units

实验耗材

分子生物学试剂

蛋白与免疫学

生化试剂

细胞及检测试剂

色谱及层析

仪器设备

实验服务

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ChemiScreen™ Membrane Prep Hu Recombinant S1P5

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