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Merck
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문서

HTS041M

Sigma-Aldrich

ChemiScreen Bradykinin B2 Membrane Preparation

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

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About This Item

UNSPSC 코드:
41106514
eCl@ss:
32161000
NACRES:
NA.41

생물학적 소스

human

Quality Level

재조합

expressed in Chem-1 cells

포장

pkg of 1 mL

제조업체/상표

ChemiScreen
Chemicon®

농도

0.5 mg/mL (per vial)

기술

ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable

NCBI 수납 번호

UniProt 수납 번호

배송 상태

dry ice

유전자 정보

human ... BDKRB2(624)

일반 설명

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.
Full length Human BDKRB2

애플리케이션

Radioligand binding assay, and GTPgammaS binding

생화학적/생리학적 작용

GPCR Class: A
Protein Target: B2
Target Sub-Family: Bradykinin

품질

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

규격

Inucbation Conditions
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

물리적 형태

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.

저장 및 안정성

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

법적 정보

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

면책조항

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 Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point (°F)

Not applicable

Flash Point (°C)

Not applicable


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