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Figure 1. Saturation binding for Oxytocin Receptor. 5.0 µg/well Oxytocin Membrane Preparation was incubated with increasing amount of [125
Figure 2. Competition binding for Oxytocin Receptor. Oxytocin Receptor Membrane Preparation (5.0 or 10µg/well) or Wild-Type Chem-1 membrane preparat...
ChemiScreen™ Recombinant Human OT Oxytocin Receptor Membrane Preparation
Description:
ChemiScreen™ Recombinant Human OT Oxytocin Receptor Membrane Preparation
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Qty/Pk:
200 units
Product Overview:
Human Oxytocin Receptor
Background Information:
Oxytocin is a cyclic 9 amino acid peptide that differs from vasopressin in 2 amino acids. Despite the close similarities in sequence, oxytocin and vasopressin have different biological activities and bind to distinct G protein-coupled receptors. The oxytocin receptor, OT, couples primarily to Gq/11 to mobilize intracellular calcium. In female reproduction, oxytocin promotes uterine contraction and lactation; oxytocin is the most commonly used drug for induction of labor, whereas an oxytocin antagonist, atosiban, is under investigation to suppress preterm labor. Oxytocin/OT interaction in the CNS also plays an important role in stress, male and female sexual response, and sociality (Gimpl and Fahrenholz, 2001). Chemicon's oxytocin 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 oxytocin receptor interactions with oxytocin. The membrane preparations exhibit a Kd of 0.53nM for [125I]-OVTA. With 5.0 ug/well Oxytocin Membrane Prep and 0.5 nM [125I]-OVTA, a greater than 6-fold signal-to-background ratio was obtained.
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.
Entrez Gene Summary:
The protein encoded by this gene belongs to the G-protein coupled receptor family and acts as a receptor for oxytocin. Its activity is mediated by G proteins which activate a phosphatidylinositol-calcium second messenger system. The oxytocin-oxytocin receptor system plays an important role in the uterus during parturition.
UniProt Summary:
FUNCTION: SwissProt: P30559 # Receptor for oxytocin. The activity of this receptor is mediated by G proteins which activate a phosphatidylinositol- calcium second messenger system.
SIZE: 389 amino acids; 42772 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P30559 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 389 amino acids; 42772 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P30559 ## Belongs to the G-protein coupled receptor 1 family.
Species:
Human
Quality Assurance:
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of Oxytocin Receptor membrane prep:
SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 4.5 pmol/mg
Kd: 0.53 nM
| 5 µg/well | 10 µg/well | |
|---|---|---|
| Signal:Background | 9 | 9 |
| Specific Binding (cpm) | 21871 | 24926 |
SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 4.5 pmol/mg
Kd: 0.53 nM
Cell Type:
Chem-1
Brand Family:
Chemicon
Incubation Conditions:
Membranes 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] OVTA (Perkin Elmer # NEX254)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
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] OVTA (Perkin Elmer # NEX254)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
Host Cells:
Chem-1
Presentation:
One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 6-fold signal:background with 125I-labeled OVTA at 0.5 nM.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Protein Target:
OT
UniProt Number:
Ligand Type:
peptide
Storage Conditions:
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Target Sub-Family:
Oxytocin
Promotional Text:
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GPCR Class:
A
Gene Symbol:
- OXTR
- OT-R
Protein or Enzyme Type:
GPCR Membrane Preparations
Product Name:
ChemiScreen™ Recombinant Human OT Oxytocin Receptor Membrane Preparation
Entrez Gene Number:
Cell Membrane Type:
GPCR