半岛bd体育手机客户端 说明
一般描述
Glucagon is a 29-amino acid peptide that stimulates glycogenolysis and gluconeogenesis in the liver to increase blood glucose. The receptor for glucagon is a class 2 (or class B) GPCR that signals through Gsto stimulate cAMP production (Mayo et al., 2003). Mice lacking the glucagon receptor have mild hypoglycemia after fasting, and exhibit hyperplasia of pancreatic α-cells (Gelling et al., 2003). Because of its role in promoting hyperglycemia, the glucagon receptor presents a potential target for treatment of diabetes. Chemicon′s glucagon receptor 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 glucagon receptor interactions and its ligands. The membrane preparations exhibit a Kd of 3.09 nM for [125I]-glucagon. With 1 nM [125I]-glucagon, 5 or 10 μg/well glucagon receptor Membrane Prep yields greater than 10-fold signal-to-background ratio.
TRANSFECTION: Full-length human GCGR cDNA encoding Glucagon Receptor
HOST CELLS: Chem-1, an adherent mammalian cell line with no endogenous glucagon receptor expression.
生化/生理作用
GPCR Class: B
Target Sub-Family: Glucagon
质量
SPECIFICATIONS:
1 unit = 5 μg
Bmaxfor [125I] glucagon binding: 16.7 pmol/mg protein
Kdfor [125I] glucagon binding: ~ 3.09 nM
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of Glucagon receptor membrane prep:
| 10 ?g/well | 5 ?g/well | |
|---|---|---|
| Signal:Background | 10.1 | 35 |
| Specific Binding (cpm) | 33315 | 36252 |
半岛bd体育手机客户端 规格
Inucbation Conditions
RECOMMENDED ASSAY CONDITIONS: Membranes are mixed with radioactive ligand and unlabeled competitor 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]-glucagon (Perkin Elmer#:NEX-207)
? 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 10-fold signal:background with [125I]-labeled glucagon at 1 nM
外形
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA; no preservatives.
Packaging method: Membrane proteins were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
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 |
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_000160.2 |
| UniProt登记号 | P47871 |
| 运输 | dry ice |
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
