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HTS083M ChemiScreen™ Recombinant Human CCK2 Cholecystokinin Receptor Membrane Preparation

Human CCK2 / CCKb GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P32239 Entrez Gene Number: NM_176875.2
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HTS083M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1CCK2 / CCKb
      Description
      Catalogue NumberHTS083M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ Recombinant Human CCK2 Cholecystokinin Receptor Membrane Preparation
      OverviewFull-length human CCKB cDNA encoding CCK2
      Background InformationCholecystokinins are a series of peptides of heterogeneous length (5 to 58 amino acids) that are derived from preprocholecystokinin and are found in gastrointestinal tissues and the central nervous system. Gastrin is a related peptide with 5 C-terminal amino acids identical to those of cholecystokinin. Two GPCRs, CCK1 (CCKA) and CCK2 (CCKB), bind to CCK and/or gastrin to mediate the biological effects of the peptides. CCK2 binds to CCK and gastrin with similar affinity, whereas CCK1 selectively binds sulfated CCK. Binding of ligands to CCK2 stimulates mobilization of intracellular calcium by activation of Gq/11. In the periphery, CCK2 is present in the stomach, where it mediates gastrin-stimulated gastric acid secretion. In the CNS, CCK2 has been implicated in anxiety, depression, schizophrenia, depression and opioid analgesia (Noble et al., 1999). Chemicon's CCK2 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 CCK2 interactions and its ligands. The membrane preparations exhibit a Kd of 0.47 nM for [125I]-cholecystokinin octapeptide (CCK-8). With 0.3 nM [125I]CCK-8, 5 or 10 μg/well CCK2 Membrane Prep yields greater than 20-fold signal-to-background ratio.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with 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 CCK2 / CCKb 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 SummaryThis gene encodes a G-protein coupled receptor for gastrin and cholecystokinin (CCK), regulatory peptides of the brain and gastrointestinal tract. This protein is a type B gastrin receptor, which has a high affinity for both sulfated and nonsulfated CCK analogs and is found principally in the central nervous system and the gastrointestinal tract. A misspliced transcript variant including an intron has been observed in cells from colorectal and pancreatic tumors.
      Gene Symbol
      • CCKBR
      • CCK-BR
      • CCK2R
      • CCK-B
      • GASR
      • CCKRB
      • CCK2-R
      GPCR ClassA
      Host CellsChem-1
      Protein TargetCCK2 / CCKb
      Target Sub-FamilyCholecystokinin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P32239 # Receptor for gastrin and cholecystokinin. The CKK-B receptors occur throughout the central nervous system where they modulate anxiety, analgesia, arousal, and neuroleptic activity. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system.
      SIZE: 447 amino acids; 48419 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P32239 ## 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C

      • Radioligand: [125I]-CCK-8 (Perkin Elmer#:NEX-203 )

      • Wash Buffer: 50 mM Tris, pH 7.4 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 20-fold signal:background with 125I labeled CCK-8 at 0.3 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 CCK2 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 27.4 27.8
      Specific Binding (cpm) 25,302 20,111


      Specifications:
      1unit = 5 μg
      Bmax for [125I] CCK-8 binding: 3.47 pmol/mg protein
      Kd for [125I] CCK-8 binding: ~0.47 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
      Catalog Number GTIN
      HTS083M 04053252515385

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      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations