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HTS023M ChemiSCREEN™ CRF1 Membrane Preparation

Human CRF1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

ChemiSCREEN™ CRF1 Membrane Preparation MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P34998 Entrez Gene Number: NM_004382.3
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HTS023M
200 assays  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1CRF1
      Description
      Catalogue NumberHTS023M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ CRF1 Membrane Preparation
      OverviewFull-length Human CRF1
      Background InformationThe CRF1 receptor is a Gs-coupled GPCR expressed in the brain and pituitary gland that binds to several neuropeptides, including corticotropin-releasing factor (CRF) and urocortin, and the amphibian peptide sauvagine (Chen et al., 1993; Dautzenberg and Hauger, 2002; Bale and Vale, 2004). CRF plays a predominant role in stress response mediated by the hypothalamic-pituitary-adrenal axis, and alterations in CRF and its receptors CRF1 and CRF2 appear to be linked to depression and anxiety (Holsboer, 1999; Bale and Vale, 2004). A number of small molecule antagonists of the CRF1 receptor have been characterized, including R121919, SC241, NBI27914, antalarmin, DMP-696, and CP 154,526. When delivered in animal models of psychiatric disorders, these antagonists display effectiveness in reducing stress-related behaviors (Kehne and De Lombaert, 2002). Chemicon's CRF1 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 CRF1 interactions with its ligands. The membrane preparations exhibit a Kd of 1 nM for [125I]-sauvagine. With 2.5 μg/well CRF1 Membrane Prep and 0.5 nM [125I]-sauvagine, a greater than 12-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 CRF1 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
      Entrez Gene Number
      Entrez Gene SummaryThe corticotropin-releasing hormone receptor binds to corticotropin-releasing hormone (MIM 122560), a potent mediator of endocrine, autonomic, behavioral, and immune responses to stress.[supplied by OMIM]
      Gene Symbol
      • CRHR1
      • CRF1
      • CRFR
      • CRHR1f
      • CRHR
      • CRFR1
      • CRF-R
      • CRH-R1h
      GPCR ClassB
      Host CellsChem-1
      Protein TargetCRF1
      Target Sub-FamilyCRF Receptor
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P34998 # This is a receptor for corticotropin releasing factor. Shows high-affinity CRF binding. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase.
      SIZE: 444 amino acids; 50719 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Predominantly expressed in the cerebellum, pituitary, cerebral cortex and olfactory lobe.
      PTM: C-terminal Ser or Thr residues may be phosphorylated.
      SIMILARITY: SwissProt: P34998 ## Belongs to the G-protein coupled receptor 2 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 (Costar 3605), 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°CRadioligand: [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 10-fold signal:background with 125I-labeled Sauvagine at 0.50 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 CRF1 membrane preparation.

      Product Usage Statements
      5 µg/well
      Signal:Background 13.2
      Specific Binding (cpm) 31785


      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 assays
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      HTS023M 04053252469596

      Documentation

      Protocols

      Title
      Assembly of a Competition Binding Curve

      ChemiSCREEN™ CRF1 Membrane Preparation SDS

      Title

      Safety Data Sheet (SDS) 

      ChemiSCREEN™ CRF1 Membrane Preparation Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN CRF1 NEUROPEPTIDE RECEPTOR - 2383396 2383396

      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
      The CRF peptide family and their receptors: yet more partners discovered
      Dautzenberg, Frank M and Hauger, Richard L
      Trends Pharmacol Sci, 23:71-7 (2002)  2002

      		11830263 11830263

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