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HTS136M ChemiScreen™ Membrane Preparation
Recombinant Human V1B Vasopressin Receptor

Human V1B GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

ChemiScreen™ Membrane Preparation
Recombinant Human V1B Vasopressin Receptor MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P47901 Entrez Gene Number: NM_000707.2
HTS136M
2 mL  200 units
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Replacement Information

Tabulka spec. kláve

SpeciesHost CellsProtein Target
Human Chem-1V1B
Description
Catalogue NumberHTS136M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiScreen™ Membrane Preparation
Recombinant Human V1B Vasopressin Receptor
OverviewHuman V1B
HOST CELLS: Chem-1, an adherent mammalian cell line without any endogenous V1B expression.
Background InformationArginine vasopressin (AVP) is a 9 amino acid peptide that functions as an antidiuretic, vasoconstrictor and neurotransmitter. The three vasopressin receptors, V1A, V1B and V2, are GPCRs; V1A and V1B couple to Gq and calcium release, whereas V2 couples to Gs (Birnbaumer, 2000). The V1B receptor is expressed prominently in the anterior pituitary, where it mediates vasopressin-induced release of ACTH (Tanoue et al., 2004). A selective antagonist of V1B has recently been developed and shown to reduce depression, anxiety, and aggression in rodents (Blanchard et al, 2005; Griebel et al., 2002). Chemicon's cloned human V1B-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant V1B expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. Chemicon's V1B 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 V1B interactions with vasopressin. The membrane preparations exhibit a Kd of 1.03 nM for [3H]-vasopressin. With 10 ug/well V1b Membrane Prep and 1.5 nM [3H]-vasopressin, a greater than 4-fold signal-to-background ratio was obtained.
References
Product Information
FormatMembranes
PresentationOne package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 4-fold signal:background with 3H-labeled vasopressin at 1.5 nM.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives. Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80&edg;C.
Quality LevelMQ100
Applications
ApplicationHuman V1B 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 SummaryThe protein encoded by this gene acts as receptor for arginine vasopressin. This receptor belongs to the subfamily of G-protein coupled receptors which includes AVPR1A, V2R and OXT receptors. Its activity is mediated by G proteins which stimulate a phosphatidylinositol-calcium second messenger system. The receptor is primarily located in the anterior pituitary, where it stimulates ACTH release. It is expressed at high levels in ACTH-secreting pituitary adenomas as well as in bronchial carcinoids responsible for the ectopic ACTH syndrome. A spliced antisense transcript of this gene has been reported but its function is not known.
Gene Symbol
  • AVPR1B
  • VPR3
  • V1bR
  • AVPR3
GPCR ClassA
Host CellsChem-1
Protein TargetV1B
Target Sub-FamilyVasopressin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P47901 # Receptor for arginine vasopressin. The activity of this receptor is mediated by G proteins which activate a phosphatidyl- inositol-calcium second messenger system.
SIZE: 424 amino acids; 46971 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P47901 ## 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 50 mM 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: [3H] vasopressin (Perkin Elmer # NET800)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceSPECIFICATIONS: 1 unit = 10 μg membrane preparation
Bmax: 2.79 pmol/mg
Kd: 1.03 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$deg;C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size2 mL
Material Package200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Katalogové číslo GTIN
HTS136M 04053252373367

Documentation

References

Reference overviewPub Med ID
AVP V1b selective antagonist SSR149415 blocks aggressive behaviors in hamsters
Blanchard, Robert J, et al
Pharmacol Biochem Behav, 80:189-94 (2005)  2004

15652395 15652395
The vasopressin V1b receptor critically regulates hypothalamic-pituitary-adrenal axis activity under both stress and resting conditions.
Tanoue, Akito, et al.
J. Clin. Invest., 113: 302-9 (2004)  2004

Zobrazit abstrakt 		14722621 14722621
Anxiolytic- and antidepressant-like effects of the non-peptide vasopressin V1b receptor antagonist, SSR149415, suggest an innovative approach for the treatment of stress-related disorders
Griebel, Guy, et al
Proc Natl Acad Sci USA, 99:6370-5 (2002)  2002

11959912 11959912
Vasopressin receptors.
Birnbaumer, M
Trends Endocrinol. Metab., 11: 406-10 (2000)  1999

Zobrazit abstrakt 		11091117 11091117

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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