ChemiScreen™ Membrane Prep Hu Recombinant PAR-2

Species	Host Cells	Protein Target
Human	Chem-1	PAR2

Species

Host Cells

Protein Target

Human

Chem-1

PAR2

Description
Catalogue Number	HTS037M
Brand Family	Chemicon®
Trade Name	ChemiScreen Chemicon
Description	ChemiSCREEN™ Membrane Preparation Recombinant Human PAR2 Protease-Activated Receptor
Overview	Human PAR2
Background Information	The protease-activated receptor family of GPCRs has a unique mechanism of activation, in which protease cleaves a prodomain to reveal a peptide sequence that functions as a tethered ligand to activate the receptor (Macfarlane et al., 2001). PAR2 is specifically activated by trypsin and mast cell tryptase, and can also be activated by free peptide analogs of the tethered ligand, such as SLIGRL and furoyl-LIGRLO (Coelho et al., 2003; Kawabata et al., 2004). PAR2 is expressed in endothelium, gastrointestinal epithelium, macrophages, eosinophils and nociceptive afferent neurons. Activation of PAR2 in these cells and tissues promotes vasodilation, inflammation, allergy, hyperalgesia and intestinal permeability. Therefore, PAR2 is regarded as an attractive therapeutic target for colitis, asthma, myocardial ischemia/reperfusion injury, and pain (Cocks et al., 1999; Hansen et al., 2005; Lindner et al., 2000; McLean et al., 2002; Vergnolle et al., 2001). Chemicon's PAR2 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 antagonists of PAR2 interactions with its ligands. The membrane preparations exhibit an EC50 of 1.9 μM for furoyl-LIGRLO in a GTPγS binding assay.

Description

Catalogue Number

HTS037M

Brand Family

Chemicon®

Trade Name

ChemiScreen
Chemicon

Description

ChemiSCREEN™ Membrane Preparation Recombinant Human PAR2 Protease-Activated Receptor

Overview

Human PAR2

Background Information

The protease-activated receptor family of GPCRs has a unique mechanism of activation, in which protease cleaves a prodomain to reveal a peptide sequence that functions as a tethered ligand to activate the receptor (Macfarlane et al., 2001). PAR2 is specifically activated by trypsin and mast cell tryptase, and can also be activated by free peptide analogs of the tethered ligand, such as SLIGRL and furoyl-LIGRLO (Coelho et al., 2003; Kawabata et al., 2004). PAR2 is expressed in endothelium, gastrointestinal epithelium, macrophages, eosinophils and nociceptive afferent neurons. Activation of PAR2 in these cells and tissues promotes vasodilation, inflammation, allergy, hyperalgesia and intestinal permeability. Therefore, PAR2 is regarded as an attractive therapeutic target for colitis, asthma, myocardial ischemia/reperfusion injury, and pain (Cocks et al., 1999; Hansen et al., 2005; Lindner et al., 2000; McLean et al., 2002; Vergnolle et al., 2001). Chemicon's PAR2 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 antagonists of PAR2 interactions with its ligands. The membrane preparations exhibit an EC50 of 1.9 μM for furoyl-LIGRLO 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 with no preservatives. Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80^oC.

Product Information

Format

Membranes

Presentation

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.

Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80^oC.

Applications
Application	Human PAR2 GPCR membrane preparation for GTPγS Binding Assays.
Key Applications	GTPγS binding
Application Notes	GTPγS Binding Assay.

Applications

Application

Human PAR2 GPCR membrane preparation for GTPγS Binding Assays.

Key Applications

GTPγS binding

Application Notes

GTPγS Binding Assay.

Biological Information
Entrez Gene Number	AY336105
Entrez Gene Summary	Coagulation factor II (thrombin) receptor-like 1 (F2RL1) is a member of the large family of 7-transmembrane-region receptors that couple to guanosine-nucleotide-binding proteins. F2RL1 is also a member of the protease-activated receptor family. It is activated by trypsin, but not by thrombin. It is activated by proteolytic cleavage of its extracellular amino terminus. The new amino terminus functions as a tethered ligand and activates the receptor. The F2RL1 gene contains two exons and is widely expressed in human tissues. The predicted protein sequence is 83% identical to the mouse receptor sequence.
Gene Symbol	F2RL1 PAR2 GPR11 PAR-2
GPCR Class	A
Host Cells	Chem-1
Protein Target	PAR2
Target Sub-Family	Protease-activated
UniProt Number	P55085
UniProt Summary	FUNCTION: SwissProt: P55085 # Receptor for trypsin and trypsin-like enzymes coupled to G proteins that stimulate phosphoinositide hydrolysis. May have a role in the regulation of vascular tone. SIZE: 397 amino acids; 44126 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. TISSUE SPECIFICITY: Widely expressed in tissues with especially high levels in pancreas, liver, kidney, small intestine, and colon. Moderate expression is detected in many organs, but none in brain or skeletal muscle. PTM: A proteolytic cleavage generates a new N-terminus that functions as a tethered ligand. SIMILARITY: SwissProt: P55085 ## Belongs to the G-protein coupled receptor 1 family.
Incubation Conditions	GTPγS 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 furoyl-LIGRLO 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 an FB filter plate (Millipore MAHF B1H) 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.

Biological Information

Entrez Gene Number

AY336105

Entrez Gene Summary

Coagulation factor II (thrombin) receptor-like 1 (F2RL1) is a member of the large family of 7-transmembrane-region receptors that couple to guanosine-nucleotide-binding proteins. F2RL1 is also a member of the protease-activated receptor family. It is activated by trypsin, but not by thrombin. It is activated by proteolytic cleavage of its extracellular amino terminus. The new amino terminus functions as a tethered ligand and activates the receptor. The F2RL1 gene contains two exons and is widely expressed in human tissues. The predicted protein sequence is 83% identical to the mouse receptor sequence.

Gene Symbol

F2RL1
PAR2
GPR11
PAR-2

GPCR Class

Host Cells

Chem-1

Protein Target

PAR2

Target Sub-Family

Protease-activated

UniProt Number

P55085

UniProt Summary

FUNCTION: SwissProt: P55085 # Receptor for trypsin and trypsin-like enzymes coupled to G proteins that stimulate phosphoinositide hydrolysis. May have a role in the regulation of vascular tone.
SIZE: 397 amino acids; 44126 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Widely expressed in tissues with especially high levels in pancreas, liver, kidney, small intestine, and colon. Moderate expression is detected in many organs, but none in brain or skeletal muscle.
PTM: A proteolytic cleavage generates a new N-terminus that functions as a tethered ligand.
SIMILARITY: SwissProt: P55085 ## Belongs to the G-protein coupled receptor 1 family.

Incubation Conditions

GTPγS 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 furoyl-LIGRLO 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 an FB filter plate (Millipore MAHF B1H) 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.

Product Usage Statements
Quality Assurance	1 unit = 5 μg EC50 for furoyl-LIGRLO: ~1.9 μM in a GTPγS binding assay
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

1 unit = 5 μg
EC50 for furoyl-LIGRLO: ~1.9 μM in a GTPγS binding assay

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	Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw.

Storage and Shipping Information

Storage Conditions

Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. 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

办公用品

ChemiScreen™ Membrane Prep Hu Recombinant PAR-2

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