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HTS041M ChemiScreen™ Bradykinin B2 Membrane Preparation

Human B2 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P30411 Entrez Gene Number: NM_000623.2
HTS041M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1B2
Description
Catalogue NumberHTS041M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiScreen™ Bradykinin B2 Membrane Preparation
OverviewFull length Human BDKRB2
Background InformationBradykinins bind and activate two GPCRs, termed B1 and B2 (gene names BDKRB1 and BDKRB2, respectively), which signal through Gq and Gi. B2 is widely expressed in a constitutive fashion, whereas B1 expression is induced during inflammation. B2 appears to mediate much of the activity of kinins in vasodilation, arthritis, edema, and nociception (Leeb-Lundberg et al., 2005). Chemicon's B2 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 B2 interactions with bradykinin. The membrane preparations exhibit a Kd of 0.18 nM for [3H]-bradykinin. With 5.0 μg/well, B2 Membrane Prep and 0.4 nM [3H]-bradykinin, a greater than 4-fold signal-to-background ratio is obtained.
References
Product Information
FormatMembranes
PresentationLiquid 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 B2 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay, and GTPgammaS binding
Biological Information
Concentration1 mL, 0.5 mg/mL per vial
Entrez Gene Number
Entrez Gene SummaryThis gene encodes a receptor for bradykinin. The 9 aa bradykinin peptide elicits many responses including vasodilation, edema, smooth muscle spasm and pain fiber stimulation. This receptor associates with G proteins that stimulate a phosphatidylinositol-calcium second messenger system. Alternate start codons result in two isoforms of the protein.
Gene Symbol
  • BDKRB2
  • BK2
  • B2R
  • DKFZp686O088
  • BRB2
  • BKR2
  • BK-2
GPCR ClassA
Host CellsChem-1
Protein TargetB2
Target Sub-FamilyBradykinin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P30411 # Receptor for bradykinin. It is associated with G proteins that activate a phosphatidylinositol-calcium second messenger system.
SIZE: 391 amino acids; 44461 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Ubiquitous. Widespread in normal smooth muscle tissue and neurons.
SIMILARITY: SwissProt: P30411 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsRECOMMENDED ASSAY 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: [3H] Bradykinin, [2,3-prolyl-3,4-3H(N)] (Perkin Elmer #NET-706)

Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, 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 4-fold signal:background with 3H-labeled bradykinin at 0.4 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceSPECIFICATIONS: 1 unit = 10 µg
Bmax: 2.73 pmol/mg
Kd: 0.18 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
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS041M 04053252284014

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