半岛bd体育手机客户端 说明
一般描述
Full-length human cDNA encoding AT1
Angiotensin II (Ang II), an octapeptide produced by cleavage of angiotensinogen by angiotensin-converting enzyme, plays a fundamental role in cardiovascular homeostasis. Two GPCRs, AT1and AT2, mediate the effects of AngII, although AT1is 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 AT1activates both Gqand Gi(De Gasparo et al., 2000). Chemicon′s AT1membrane 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 AT1interactions 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 ?g/well AT1Membrane Prep yields greater than 50-fold signal-to-background ratio.
应用
Radioligand binding assay and GTPγS binding.
生化/生理作用
GPCR Class: A
Target Sub-Family: Angiotensin
Protein Target: AT1
质量
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of AT
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SPECIFICATIONS: 1 unit = 5 ?g
Bmax for [125I] Angiotensin II binding: 5.53 pmol/mg protein;
Kd for [125I] Angiotensin II binding: ~ 0.36 nM
半岛bd体育手机客户端 规格
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 with125I labeled Sar1-Ile8-Angiotensin II at 0.3 nM
外形
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.
储存及稳定性
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
法律信息
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免责声明
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.
基本信息
| eCl@ss | 32161000 |
| NACRES | NA.84 |
半岛bd体育手机客户端 性质
| 质量水平 | 100 |
| 生物来源 | human |
| 重组 | expressed in Chem-1 cells |
| manufacturer/tradename | ChemiScreen Chemicon? |
| technique(s) | ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable |
| NCBI登记号 | NM_000685.4 NM_004835.3 NM_009585.3 NM_031850.2 NM_032049.2 |
| UniProt登记号 | P30556 |
| 运输 | dry ice |
| Gene Information | human ... AGTR1(185) |
安全信息
| 储存分类代码 | 12 - Non Combustible Liquids |
| WGK | WGK 2 |
| 闪点(F) | Not applicable |
| 闪点(C) | Not applicable |
Sigma-Aldrich
m.cnreagent.com
