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HTS074M ChemiSCREEN™ Human PK1 Prokineticin Receptor Membrane Preparation

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HTS074M
200 units  
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Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1PK1 / PKR1
Description
Catalogue NumberHTS074M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Human PK1 Prokineticin Receptor Membrane Preparation
OverviewFull-length human GPR73 cDNA, encoding PK1
Background InformationProkineticins, also known as endocrine gland vascular endothelial growth factors (EG-VEGF), are two ~10 kD secreted proteins originally described to mediate angiogenesis and gastrointestinal smooth muscle contraction (Li et al., 2001; LeCouter et al., 2003). Subsequently, prokineticins have been found to mediate central nervous system functions including circadian rhythms and olfactory bulb development (Cheng et al., 2002; Ng et al., 2005). Two Gq-coupled receptors, PK1 and PK2 (also known as GPR73a and GPR73b), mediate cellular responses to prokineticins (Lin et al., 2002). Chemicon's PK1 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 PK1 interactions and its ligands. The membrane preparations exhibit a Kd of 0.11 nM for [125I]-Mamba Intestinal Toxin-1 (MIT-1). With 0.2 nM [125I]-MIT-1, 2.5 µg/well and 5 µg/well PK1 Membrane Prep typically yield greater than 5-fold signal-to-background ratio.
References
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 0.5 mg/ml in 1 mL packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman PK1 / PKR1 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
  • PROKR1
  • PK-R1
  • PKR1
  • GPR73
  • GPR73a
  • ZAQ
GPCR ClassA
Host CellsChem-1
Protein TargetPK1 / PKR1
Target Sub-FamilyProkineticin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q8TCW9 # Receptor for prokineticin 1. Exclusively coupled to the G(q) subclass of heteromeric G proteins. Activation leads to mobilization of calcium, stimulation of phosphoinositide turnover and activation of p44/p42 mitogen-activated protein kinase.
SIZE: 393 amino acids; 44770 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed in the stomach, throughout the small intestine, colon, rectum, thyroid gland, pituitary gland, salivary gland, adrenal gland, testis, ovary, brain, spleen, prostate and pancreas.
SIMILARITY: SwissProt: Q8TCW9 ## 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]-MIT-1 (Perkin Elmer#:NEX-410 )
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I-labeled MIT-1 at 0.2 nM
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 PK1 membrane prep:
Product Usage Statements
5 µg/well 2.5 µg/well
Signal:Background 6 6.4
Specific Binding (cpm) 3166 1798


SPECIFICATIONS: 1 unit = 5 µg
Bmax for [125I] MIT-1 binding: 0.58 pmol/mg protein;
Kd for [125I] MIT-1 binding: ~ 0.11 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
Catalogue Number GTIN
HTS074M 04053252326592

Documentation

ChemiSCREEN™ Human PK1 Prokineticin Receptor Membrane Preparation SDS

Title

Safety Data Sheet (SDS) 

References

Reference overviewPub Med ID
Dependence of olfactory bulb neurogenesis on prokineticin 2 signaling
Ng, Kwan L, et al
Science, 308:1923-7 (2005)  2005

15976302 15976302
Prokineticin 2 transmits the behavioural circadian rhythm of the suprachiasmatic nucleus.
Cheng, Michelle Y, et al.
Nature, 417: 405-10 (2002)  2002

Show Abstract
12024206 12024206
Endocrine gland-derived VEGF and the emerging hypothesis of organ-specific regulation of angiogenesis.
LeCouter, Jennifer, et al.
Nat. Med., 8: 913-7 (2002)  2002

Show Abstract
12205443 12205443

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Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations