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HTS024M ChemiSCREEN™ Membrane Preparation Recombinant Human CRF2 Neuropeptide Receptor

Human CRF2 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

ChemiSCREEN™ Membrane Preparation Recombinant Human CRF2 Neuropeptide Receptor MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: Q13324 Entrez Gene Number: NM_001883.3
HTS024M
200 units  
Purchase on Sigma-Aldrich

Przegląd

Replacement Information

Tabela kluczowych gatunków

SpeciesHost CellsProtein Target
Human Chem-1CRF2
Description
Catalogue NumberHTS024M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human CRF2 Neuropeptide Receptor
OverviewFull-length human CRHR2 cDNA encoding CRF2
Background InformationThe corticotropin-releasing factor receptors, CRF1 and CRF2, are Gs-coupled GPCRs expressed in the brain, blood vessels and intestine that bind to several neuropeptides, including corticotropin-releasing factor (CRF) and urocortin, and the amphibian peptide sauvagine (Lovenberg et al., 1995; Bale and Vale, 2004). In addition, two peptides, urocortin II (Ucn II) and urocortin III (Ucn III), bind selectively and with high affinity to CRF2 (Lewis et al., 2001). The CRF peptides and their receptors play important roles in stress mediated by the hypothalamic-pituitary-adrenal axis in animal models, and possibly in depression and anxiety in humans, although the contributions of CRF1 and CRF2 appear to be distinct (Bale and Vale, 2004; Risbrough et al., 2004) In addition, CRF1 and CRF2 differentially alter feeding behavior, gastric motility and vascular tone (Zorrilla et al., 2004; Martinez et al., 2004; Wiley and Davenport, 2004). Millipore's CRF2 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 CRF2 interactions with its ligands. The membrane preparations exhibit a Kd of 0.65 nM for [125I]-sauvagine. With 5 μg/well CRF2 Membrane Prep and 0.35 nM [125I]-sauvagine, a greater than 20-fold signal-to-background ratio is obtained.
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 CRF2 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay, and GTPgammaS binding.
Biological Information
Concentration0.5 mg/mL
Entrez Gene Number
Gene Symbol
  • CRHR2
  • CRH2R
  • CRF2
  • CRFR2
  • CRF2R
GPCR ClassB
Host CellsChem-1
Protein TargetCRF2
Target Sub-FamilyCRF Receptor
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q13324 # This is a receptor for corticotropin releasing factor. Shows high-affinity CRF binding. Also binds to urocortin I, II and III. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase.
SIZE: 411 amino acids; 47688 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: Q13324 ## Belongs to the G-protein coupled receptor 2 family.
Incubation ConditionsRECOMMENDED ASSAY CONDITIONS: Membranes 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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) 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] Sauvagine (Perkin Elmer#:NEX306)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

One vial contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 20-fold signal:background with 125I-labeled Sauvagine at 0.35 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. Signal:background and specific binding values obtained in a competition binding assay with CRF2 membrane preparationn.
Product Usage Statements
5 µg/well
Signal:Background 64.6
Specific Binding (cpm) 38718


SPECIFICATIONS: 1 unit = 5 µg
Bmax: 9.04 pmol/mg protein
Kd : 0.65 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
Numer katalogowy GTIN
HTS024M 04053252333170

Documentation

Protocols

Title
Assembly of a Competition Binding Curve

ChemiSCREEN™ Membrane Preparation Recombinant Human CRF2 Neuropeptide Receptor MSDS

Title

Safety Data Sheet (SDS) 

ChemiSCREEN™ Membrane Preparation Recombinant Human CRF2 Neuropeptide Receptor Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CRF2 CORTICOTROPIN RELEASING FACTOR RECEPTOR - 2203660 2203660
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CRF2 CORTICOTROPIN RELEASING FACTOR RECEPTOR - 2250856 2250856
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CRF2 CORTICOTROPIN RELEASING FACTOR RECEPTOR - 2326012 2326012

References

Reference overviewPub Med ID
CRF and CRF receptors: role in stress responsivity and other behaviors
Bale, Tracy L and Vale, Wylie W
Annu Rev Pharmacol Toxicol, 44:525-57 (2004)  2004

14744257 14744257

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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