HTS123M

ChemiScreen Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation

Human BB1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

• 分類程式碼代碼: 41106514
• eCl@ss: 32161000
• NACRES: NA.84

屬性

• 生物源: human
• 品質等級: 100
• 重組細胞: expressed in Chem-1 cells
• 製造商／商標名: ChemiScreen; Chemicon^®
• 技術: ligand binding assay: suitable (GTPγS); radioligand binding assay (RLBA): suitable
• NCBI登錄號: NM_002511.2
• UniProt登錄號: P28336
• 運輸包裝: dry ice
• 基因資訊: human ... NMBR(4829)

描述

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 (NMB). 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; BB2 binds to GRP with 50-300-fold greater affinity than to NMB, whereas BB1 binds to NMB with 10-800-fold greater affinity than to GRP (Tokita et al., 2004). Binding of ligand to BB1 activates Gq to increase intracellular calcium concentrations. The CNS is a major site of NMB and BB1 expression, and BB₁ appears to be involved in thermoregulation (Ohki-Hamazaki et al., 2005). Chemicon′s BB₁ 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 BB1 interactions with neuromedin B. The membrane preparations exhibit a Kd of 0.35-0.43 nM for [¹²⁵I]-Bombesin. With 5 µg/well BB₁ Membrane Prep and 0.3 nM [¹²⁵I]-bombesin, a greater than 40-fold signal-to-background ratio was obtained.

Full-length human NMBR cDNA encoding BB1

應用

Radioligand binding assay, and GTPγS binding.

生化／生理作用

GPCR Class: A

Protein Target: BB1

Target Sub-Family: Bombesin

包裝

1 mL per vial

品質

Signal:background and specific binding values obtained in a competition binding assay with varying amounts of BB₁ membrane prep:

	10 µg/well	5 µg/well
Signal:Background	82.5	74.8
Specific Binding (cpm)	27377	22443

SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 3.1 pmol/mg
Kd: 0.39 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 Hepes, pH 7.4, 5 mM MgCl₂, 1 mM CaCl₂, 0.2% BSA, filtered and stored at 4°C
Radioligand: [¹²⁵I] bombesin (Perkin Elmer # NEX258)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.

外觀

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 40-fold signal:background with ¹²⁵I-labeled bombesin at 0.3 nM.

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein were adjusted to the indicated concentration in 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

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.

安全資訊

• 儲存類別代碼: 10 - Combustible liquids
• 水污染物質分類（WGK）: WGK 2