ChemiScreen H1 Membrane Preparation

半岛bd体育手机客户端 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 ?g/well
Signal:Background	6.6
Specific Binding (cpm)	925

SPECIFICATIONS: 1 unit = 10 ?g for most lots. Specific activity (units/?g) 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 ?g

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 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

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-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