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ChemiScreen™ Membrane Prep Hu Recombinant CRF2

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货号：HTS024M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-1 CRF2
Description
Catalogue Number HTS024M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiSCREEN™ Membrane Preparation Recombinant Human CRF2 Neuropeptide Receptor
Overview Full-length human CRHR2 cDNA encoding CRF2
Background Information The 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
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 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
Applications
Application Human CRF2 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application Notes Radioligand binding assay, and GTPgammaS binding.
Biological Information
Concentration 0.5 mg/mL
Entrez Gene Number
Gene Symbol
  • CRHR2
  • CRH2R
  • CRF2
  • CRFR2
  • CRF2R
GPCR Class B
Host Cells Chem-1
Protein Target CRF2
Target Sub-Family CRF Receptor
UniProt Number
UniProt Summary FUNCTION: 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 Conditions RECOMMENDED 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
2 Table 1. Signal:background and specific binding values obtained in a competition binding assay with CRF 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 Conditions Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size 200 units
Transport Information
Supplemental Information
Specifications

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