HTS090M
ChemiScreen Recombinant Human OT Oxytocin Receptor Membrane Preparation
Human OT GPCR membrane prepation for Radioligand binding Assays.
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About This Item
UNSPSC Code:
41106514
eCl@ss:
32161000
NACRES:
NA.84
biological source
human
recombinant
expressed in Chem-1 cells
manufacturer/tradename
ChemiScreen
Chemicon®
technique(s)
radioligand binding assay (RLBA): suitable
NCBI accession no.
UniProt accession no.
shipped in
dry ice
Gene Information
human ... OXTR(5021)
General description
Human Oxytocin Receptor
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.
Biochem/physiol Actions
GPCR Class: A
Protein Target: OT
Target Sub-Family: Oxytocin
Quality
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
Specifications
Inucbation 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.
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.
Physical form
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.
Storage and Stability
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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 Class
12 - Non Combustible Liquids
wgk_germany
WGK 2
flash_point_f
Not applicable
flash_point_c
Not applicable
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G Gimpl et al.
Physiological reviews, 81(2), 629-683 (2001-03-29)
The neurohypophysial peptide oxytocin (OT) and OT-like hormones facilitate reproduction in all vertebrates at several levels. The major site of OT gene expression is the magnocellular neurons of the hypothalamic paraventricular and supraoptic nuclei. In response to a variety of
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