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HTS059M ChemiScreen™ V1A Vasopressin Membrane Preparation

HTS059M
200 units  1 mL, 2 mg/mL
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1V1A
Description
Catalogue NumberHTS059M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiScreen™ V1A Vasopressin Membrane Preparation
OverviewHuman AVPR1 cDNA encoding V1A
Materials Required but Not DeliveredMembranes 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 Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C Radioligand: [125I] vasopressin V1A antagonist (Perkin Elmer catalog # NEX310)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
One 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 125I-labeled vasopressin V1A antagonist at 0.1 nM
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. V1 receptors regulate fluid homeostasis by stimulating contraction of glomerular mesangial cells and renal vascular smooth muscle cells (Birnbaumer, 2000). Chemicon's V1A 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 V1A interactions with its ligands. The membrane preparations exhibit a Kd of 55 pM for [125I]-vasopressin antagonist ([125I]-phenylacetyl-D-Tyr(Me)-Phe-Gln-Asn-Arg -Pro-Arg-Tyr-NH2). With 10 µg/well V1A Membrane Prep and 0.1 nM [125]-vasopressin V1A antagonist, greater than 12-fold signal-to-background ratio were 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 was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman V1A 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
Concentration0.5 mg/mL
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 AVPR1B, V2R and OXT receptors. Its activity is mediated by G proteins which stimulate a phosphatidylinositol-calcium second messenger system. The receptor mediates cell contraction and proliferation, platelet aggregation, release of coagulation factor and glycogenolysis.
Gene Symbol
  • AVPR1A
  • AVPR1
  • V1aR
GPCR ClassA
Host CellsChem-1
Protein TargetV1A
Target Sub-FamilyVasopressin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P37288 # Receptor for arginine vasopressin. The activity of this receptor is mediated by G proteins which activate a phosphatidyl- inositol-calcium second messenger system. Has been involved in social behaviors, including affiliation and attachment.
SIZE: 418 amino acids; 46800 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P37288 ## Belongs to the G-protein coupled receptor 1 family.
MISCELLANEOUS: Differences in regional receptor expression in the brain as well as differences in social behavior may result from a highly variable repetitive sequence in the 5' flanking region of AVPR1A. One such allelic variant has been linked to autism.
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 V1A membrane prep:
Product Usage Statements
10 µg/well
Signal:Background 24.4
Specific Binding (cpm) 11827


SPECIFICATIONS:
1 unit = 10 µg
Bmax: 1.76 pmol/mg
Kd: 55 pM
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
Material Package1 mL, 2 mg/mL
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS059M 04053252506598

Documentation

ChemiScreen™ V1A Vasopressin Membrane Preparation SDS

Title

Safety Data Sheet (SDS) 

References

Reference overviewPub Med ID
Vasopressin receptors.
Birnbaumer, M
Trends Endocrinol. Metab., 11: 406-10 (2000)  2000

Show Abstract
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