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HTS149M ChemiSCREEN™ Membrane Preparation Recombinant Human mGlu6 Metabotropic Glutamate Receptor

Human mGlu6 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: O15303 Entrez Gene Number: NM_000843.3
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HTS149M
200 units  
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      Übersicht

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1mGlu6
      Description
      Catalogue NumberHTS149M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human mGlu6 Metabotropic Glutamate Receptor
      Overviewhuman GRM6 cDNA
      Background InformationGlutamate is a main excitatory neurotransmitter in the central nervous system, and it plays a role in learning, memory and neurotoxicity. The biological actions of glutamate are mediated by ionotropic and metabotropic glutamate receptors, which are ion channels and GPCRs respectively. Metabotropic glutamate receptors (mGluRs) are members of the class 3 G-protein coupled receptor family, which are characterized by a large extracellular domain. They are further classified into group I, II, and III mGluRs on the basis of their sequence identity, pharmacology, and signal transduction mechanism. Group I (mGlu1 and mGlu5) couple to the phospholipase C pathway through Gαq, whereas group II (mGlu2 and mGlu3) and group III (mGlu4, mGlu6, mGlu7, and mGlu8) negatively couple to the adenylyl cyclase pathway though Gαi (Conn and Pin, 1997). mGlu6 has a restricted localization to the dendrites of ON bipolar cells that synapse with retinal rods and cones. Targeted deletion of mGlu6 results in deficits in ON responses but unchanged OFF responses to light (Masu et al., 1995). In addition, mutations in GRM6 in humans are responsible for some forms of autosomal recessive congenital stationary night blindness (Dryja et al., 2005; Zeitz et al., 2005). Chemicon's mGlu6 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 mGlu6 interactions with its ligands. The cell line exhibits a calcium response with EC50s of 6.2 µM, 23.4 µM, and 52.7 µM for L-AP4, (RS)PPG, and L-glutamate. The membrane preparations exhibit EC50s of of 4.0 µM, 24.1 µM, and 49.3 µM for L-AP4, (RS)PPG, and L-glutamate, respectively, in a GTPγS binding assay.
      References
      Product Information
      FormatMembranes
      PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
      Packaging method: Membrane protein was adjusted to 1 mg/ml in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman mGlu6 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 SummaryL-glutamate is the major excitatory neurotransmitter in the central nervous system and activates both ionotropic and metabotropic glutamate receptors. Glutamatergic neurotransmission is involved in most aspects of normal brain function and can be perturbed in many neuropathologic conditions. The metabotropic glutamate receptors are a family of G protein-coupled receptors, that have been divided into 3 groups on the basis of sequence homology, putative signal transduction mechanisms, and pharmacologic properties. Group I includes GRM1 and GRM5 and these receptors have been shown to activate phospholipase C. Group II includes GRM2 and GRM3 while Group III includes GRM4, GRM6, GRM7 and GRM8. Group II and III receptors are linked to the inhibition of the cyclic AMP cascade but differ in their agonist selectivities.
      Gene Symbol
      • GRM6
      • DKFZp686H1993
      • GPRC1F
      • mGluR6
      • MGLUR6
      • CSNB1B
      • mGlu6
      GPCR ClassC
      Host CellsChem-1
      Protein TargetmGlu6
      Target Sub-FamilyGlutamate (metabotropic)
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: O15303 # Receptor for glutamate. The activity of this receptor is mediated by a G-protein that inhibits adenylate cyclase activity.
      SIZE: 877 amino acids; 95437 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      DISEASE: SwissProt: O15303 # Defects in GRM6 are the cause of congenital stationary night blindness type 1B (CSNB1B) [MIM:257270]. This disorder consits of a previously unrecognized, autosomal recessive form of congenital night blindness associated with a negative electroretinogram waveform. Patients are night blind from an early age, and when maximally dark-adapted, they could perceive lights only with an intensity equal to or slightly dimmer than that normally detected by the cone system. ERGs in response to single brief flashes of light have clearly detectable a-waves, which are derived from photoreceptors, and greatly reduced b-waves, which are derived from the second-order inner retinal neurons. ERGs in response to sawtooth flickering light indicate a markedly reduced ON response and a nearly normal OFF response. There is no subjective delay in the perception of suddenly appearing white vs black objects on a gray background.
      SIMILARITY: SwissProt: O15303 ## Belongs to the G-protein coupled receptor 3 family.
      Incubation ConditionsMembranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [35S]-GTPγS (final concentration of 0.3 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl2/0.5 µM GDP in a nonbinding 96-well plate. Unlabeled DCG IV, (2R4R) APDC, and glutamate are added to the final concentration indicated in Figure 1 (final volume 100 µL), and incubated for 30 min at 30°C. The binding reaction is transferred to a GF/B filter plate (Millipore MAHF B1H) previously prewetted with water, and washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4. The plate is dried and counted.

      One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific L-AP4, (RS)PPG, or glutamate -stimulated [35S]-GTPγS binding.
      The mGlu6 membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEC50 in GTPγS binding assay by Glutamate: ~ 49.3 μM
      EC50 in GTPγS binding assay by (RS)PPG: ~ 24.1 μM
      EC50 in GTPγS binding assay by L-AP4: ~ 4.04 μM
      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
      Bestellnummer GTIN
      HTS149M 04053252734236

      Documentation

      ChemiSCREEN™ Membrane Preparation Recombinant Human mGlu6 Metabotropic Glutamate Receptor SDB

      Titel

      Sicherheitsdatenblatt (SDB) 

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      Kategorien

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