ChemiScreen™ Bradykinin B₂ Membrane Preparation

ChemiScreen™ Bradykinin B₂ Membrane Preparation

• ChemiScreen
• Chemicon (Millipore)

200 units

Full length Human BDKRB2

Bradykinins 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.

Radioligand binding assay, and GTPgammaS binding

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.

This 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.

FUNCTION: 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.

Human

Chemicon

B2

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.

peptide

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Bradykinin

A

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• BDKRB2
• BK2
• B2R
• DKFZp686O088
• BRB2
• BKR2
• BK-2

GPCR Membrane Preparations

ChemiScreen™ Bradykinin B₂ Membrane Preparation

1 mL, 0.5 mg/mL per vial

SPECIFICATIONS: 1 unit = 10 µg
Bmax: 2.73 pmol/mg
Kd: 0.18 nM

Chem-1

RECOMMENDED 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

Chem-1

P30411

NM_000623.2

GPCR