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HTS123M ChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation

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

ChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P28336 Entrez Gene Number: NM_002511.2
HTS123M
200 units  1 mL per vial
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1BB1
Description
Catalogue NumberHTS123M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation
OverviewFull-length human NMBR cDNA encoding BB1
Background InformationBombesin, 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.
References
Product Information
FormatMembranes
PresentationOne 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.
Quality LevelMQ100
Applications
ApplicationHuman BB1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay, and GTPγS binding.
Biological Information
Entrez Gene Number
Entrez Gene SummaryNeuromedin B receptor binds neuromedin B, a potent mitogen and growth factor for normal and neoplastic lung and for gastrointestinal epithelial tissue.
Gene Symbol
  • NMBR
  • NMB-R
GPCR ClassA
Host CellsChem-1
Protein TargetBB1
Target Sub-FamilyBombesin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P28336 # Receptor for neuromedin-B.
SIZE: 390 amino acids; 43453 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P28336 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsMembranes 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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of BB1 membrane prep:
Product Usage Statements
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 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.
Storage and Shipping Information
Storage ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 units
Material Package1 mL per vial
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS123M 04053252624124

Documentation

ChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation MSDS

Title

Safety Data Sheet (SDS) 

ChemiScreen™ Recombinant Human BB1 Bombesin/Neuromedin B Receptor Membrane Preparation Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN BB1 BOMBESIN/NEUROMEDIN B RECEPTOR - 2037872 2037872

References

Reference overviewPub Med ID
Development and function of bombesin-like peptides and their receptors
Ohki-Hamazaki, Hiroko, et al
Int J Dev Biol, 49:293-300 (2005)  2004

15906244 15906244
Molecular basis of the selectivity of gastrin-releasing peptide receptor for gastrin-releasing peptide
Tokita, Kenji, et al
Mol Pharmacol, 61:1435-43 (2002)  2002

12021405 12021405

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