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一般說明
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
應用
Human BB2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding.
生化／生理作用
GPCR Class: A
Protein Target: BB2
Target Sub-Family: Bombesin
品質
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
規格
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
外觀
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.
儲存和穩定性
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
法律資訊
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.
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.
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.
儲存類別代碼
12 - Non Combustible Liquids
水污染物質分類（WGK）
WGK 2
閃點（°F）
Not applicable
閃點（°C）
Not applicable
分析證明 (COA)
輸入產品批次/批號來搜索 分析證明 (COA)。在產品’s標籤上找到批次和批號，寫有 ‘Lot’或‘Batch’.。
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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