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HTS057M ChemiScreen™ Recombinant Human PRP/GPR10 Prolactin-Releasing Peptide Receptor Membrane Preparation

Human PRP / PrRP / GPR10 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

Host Cell: Chem-1 Uni Prot Number: P49683 Entrez Gene Number: NM_004248.2
HTS057M
200 units  1 mL, 1 mg/mL
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Specifications Table

SpeciesHost CellsProtein Target
Human Chem-1PRP / PrRP / GPR10
Description
Catalogue NumberHTS057M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiScreen™ Recombinant Human PRP/GPR10 Prolactin-Releasing Peptide Receptor Membrane Preparation
OverviewHuman Full-length human GPR10 cDNA encoding PRP
Background InformationPRP, also known as GPR10 or hGR3, is a Gq-coupled receptor for prolactin-releasing peptide that is expressed in the pituitary (Hinuma et al., 1998). Genetic studies in rodents indicate that lack of GPR10 leads to hyperphagia, obesity and dyslipidemia (Gu et al., 2004; Watanabe et al., 2005). In humans, genetic variations in GPR10 are associated with lowered blood pressure (Bhattacharyya et al., 2003). Chemicon's GPR10/PRP 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 GPR10/PRP interactions with prolactin-releasing peptide. The membrane preparations exhibit a Kd of 0.59-0.66 nM for [125I]-PRP-20. With 5 µg/well PRP Membrane Prep and 0.25 nM [125I]-PRP-20, a greater than 30-fold signal-to-background ratio was obtained.
References
Product Information
FormatMembranes
PresentationOne package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 30-fold signal:background with 125I-labeled PRP-20 at 0.25 nM.

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°C.
Quality LevelMQ100
Applications
ApplicationHuman PRP / PrRP / GPR10 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 SummaryPRLHR is a 7-transmembrane domain receptor for prolactin-releasing hormone (PRLH; MIM 602663) that is highly expressed in anterior pituitary (Ozawa et al., 2002 [PubMed 11923475]).[supplied by OMIM]
Gene Symbol
  • PRLHR
  • GR3
  • GPR10
  • MGC126539
  • PrRPR
  • hGR3
  • MGC126541
GPCR ClassA
Host CellsChem-1
Protein TargetPRP / PrRP / GPR10
Target Sub-FamilyProlactin-releasing peptide
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P49683 # Receptor for prolactin-releasing peptide (PrRP). Implicated in lactation, regulation of food intake and pain-signal processing.
SIZE: 370 amino acids; 41107 Da
SUBUNIT: Interacts through its C-terminal region with the PDZ domain-containing proteins GRIP1, GRIP2 and PICK1. Interacts with PDZ domains 4 and 5 of GRIP1 and with the PDZ domain of PICK1.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Only detected in the pituitary gland and in all cell types of pituitary adenomas.
SIMILARITY: SwissProt: P49683 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsMembranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
Radioligand: [125I] PRP-20 (Perkin Elmer # NEX384)
Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of PRP membrane prep:
Product Usage Statements
10 µg/well 5 µg/well
Signal:Background 46.0 51.6
Specific Binding (cpm) 15756 14148


SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 8.9 pmol/mg
Kd: 0.62 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
Material Package1 mL, 1 mg/mL
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
HTS057M 04053252338083

Documentation

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