产品说明
一般描述
Histamine exerts its biological effects through a family of four GPCRs. Stimulation of the H1 histamine receptor causes smooth muscle contraction, induces vascular permeability, and stimulates catecholamine release from the adrenal medulla. H1 is also widely expressed in brain, and activation of H1 excites several types of neurons by blocking potassium conductance (Hill et al., 1997).
Chemicon′s H1 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 H1 interactions and its ligands. The membrane preparations exhibit a Kd of 4.5 nM for [3H]-Pyrilamine. With 3 nM [3H]-Pyrilamine, 5 mg/well and 10 mg/well H1 Membrane Prep typically yield greater than 3 and 6-fold signal-to-background ratios, respectively.
Full-length human HRH1 cDNA
应用
Radioligand binding assay, and GTPγS binding.
生化/生理作用
Protein Target: H1
Target Sub-Family: Histamine
GPCR Class: A
质量
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of TP membrane prep:
| 10 µg/well | |
|---|---|
| Signal:Background | 6.6 |
| Specific Binding (cpm) | 925 |
SPECIFICATIONS: 1 unit = 10 µg for most lots. Specific activity (units/µg) may vary between lots.
Bmax: 8.3 pmol/mg
Kd: 4.5 nM
Note: Lot number MM1357015 contains 2 mL at 2 mg/mL, for which 1 unit = 20 µg
产品规格
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 50 mM 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C
Radioligand: [3H] Pyrilamine (Perkin Elmer#:NET594250UC )
Wash Buffer: 50 mM Tris, pH 7.4 filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 3-fold signal:background with 3H-labeled Pyrilamine at 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 -80°C.
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 |
产品性质
| 质量水平 | 100 |
| 生物来源 | human |
| 重组 | expressed in Chem-3 cells |
| 包装 | pkg of 2 mL |
| manufacturer/tradename | ChemiScreen Chemicon® |
| 浓度 | 0.5 mg/mL |
| technique(s) | ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable |
| NCBI登记号 | NM_000861.3 NM_001098211.1 NM_001098212.1 NM_001098213.1 |
| UniProt登记号 | P35367 |
| 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 |
