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Key Documents

HTS064M

Sigma-Aldrich

ChemiSCREEN Human Angiotensin II Receptor, Type I Membrane Preparation

Human AT1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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About This Item

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

生物源

human

品質等級

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

UniProt登錄號

運輸包裝

dry ice

基因資訊

human ... AGTR1(185)

一般說明

Angiotensin II (Ang II), an octapeptide produced by cleavage of angiotensinogen by angiotensin-converting enzyme, plays a fundamental role in cardiovascular homeostasis. Two GPCRs, AT1 and AT2, mediate the effects of AngII, although AT1 is primarily responsible for the effects of Ang II on renal function and development, thirst, electrolyte and water balance, and arterial blood pressure. Binding of Ang II to AT1 activates both Gq and Gi (De Gasparo et al., 2000). Chemicon′s AT1 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 AT1 interactions and its ligands. The membrane preparations exhibit a Kd of 0.36 nM for [125I]-Sar1-Ile8-Angiotensin II. With 0.3 nM [125I]-Sar1-Ile8-Angiotensin II, 5 µg/well AT1 Membrane Prep yields greater than 50-fold signal-to-background ratio.
Full-length human cDNA encoding AT1

應用

Radioligand binding assay and GTPγS binding.

生化/生理作用

GPCR Class: A
Protein Target: AT1
Target Sub-Family: Angiotensin

品質

Signal:background and specific binding values obtained in a competition binding assay with varying amounts of ATaub1 membrane prep:
tabletrth align="left"/tdth align="center"10 µg/well/tdth align="center"5 µg/well /td/trtrtd align="left" Signal:Background/tdtd align="center" 115.5/tdtd align="center" 92.8/td/trtrtd align="left" Specific Binding (cpm) /tdtd align="center"46,458/tdtd align="center"43,007 /td/tr/table

SPECIFICATIONS: 1 unit = 5 µg
Bmax for [125I] Angiotensin II binding: 5.53 pmol/mg protein;
Kd for [125I] Angiotensin II binding: ~ 0.36 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]-Sar1-Ile8-Angiotensin II. (Perkin Elmer#:NEX-248 )
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM 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 50-fold signal:background with 125I labeled Sar1-Ile8-Angiotensin II at 0.3 nM

外觀

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.
Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80oC.

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.

儲存類別代碼

12 - Non Combustible Liquids

水污染物質分類(WGK)

WGK 2

閃點(°F)

Not applicable

閃點(°C)

Not applicable


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