HTS084M
ChemiScreen BB2 Bombesin Receptor Membrane Preparation
Human BB2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
Code UNSPSC :
41106514
eCl@ss :
32161000
Nomenclature NACRES :
NA.84
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Description générale
Bombesin, a bioactive peptide first identified in amphibian skin, is related to two mammalian peptides, gastrin-releasing peptide (GRP) and neuromedin B. A family of 3 GPCRs, including NMB-R (BB1), GRP-R (BB2) and BRS-3 (BB3), mediate the biological effects of the peptides. The receptors differ in their affinities for the peptides. Binding of ligand to BB2 activates Gq to increase intracellular calcium concentrations. GRP stimulates release of gastrin from endocrine cells and stimulates smooth muscle activity in the gastrointestinal tract. In addition, binding of GRP to BB2 stimulates proliferation of a variety of cell types, and has been implicated in the progression of small cell lung cancer and other malignancies. The CNS is also a major site of GRP expression, and GRP and BB2 are involved in the circadian clock, conditioned fear, and food intake (Ohki-Hamazaki et al., 2005). Chemicon′s BB2 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 BB2 interactions and its ligands. The membrane preparations exhibit a Kd of 0.114 nM for [125I]-Tyr4-Bombesin. With 0.3nM [125I]-Tyr4-Bombesin, 5 µg/well or 10 µg/well BB2 Membrane Prep typically yield greater than 20-fold signal-to-background ratio.
Full-length human GRPR cDNA encoding BB2
Application
Human BB2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding.
Actions biochimiques/physiologiques
GPCR Class: A
Protein Target: BB2
Target Sub-Family: Bombesin
Qualité
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of BB2 membrane prep:
SPECIFICATION: Bmax: 1.28 pmol/mg; Kd: 0.114 nM
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 24 | 23 |
| Specific Binding (cpm) | 19,260 | 12,610 |
SPECIFICATION: Bmax: 1.28 pmol/mg; Kd: 0.114 nM
Caractéristiques
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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C
Radioligand: [125I]-Tyr4-Bombesin (Perkin Elmer#:NEX-258 )
Wash Buffer: 50 mM Tris, pH 7.4 filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I labeled Bombesin at 0.3 nM
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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C
Radioligand: [125I]-Tyr4-Bombesin (Perkin Elmer#:NEX-258 )
Wash Buffer: 50 mM Tris, pH 7.4 filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I labeled Bombesin at 0.3 nM
Forme physique
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.
Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
Stockage et stabilité
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Informations légales
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Clause de non-responsabilité
Ce produit, destiné à la recherche scientifique, est soumis à une réglementation spécifique en France, y compris pour les activités d′importation et d′exportation (Article L 1211-1 alinéa 2 du Code de la Santé Publique). L′acheteur (c′est-à-dire l′utilisateur FINAL) est tenu d′obtenir une autorisation d′importation auprès du ministère français de la recherche, mentionné à l′article L1245-5-1 II du Code de la Santé Publique. En commandant ce produit, vous confirmez détenir l′autorisation d′importation requise.
Code de la classe de stockage
12 - Non Combustible Liquids
Classe de danger pour l'eau (WGK)
WGK 2
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
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Development and function of bombesin-like peptides and their receptors
Ohki-Hamazaki, Hiroko, et al
International Journal of Developmental Biology, 49, 293-300 (2005)
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