ChemiScreen Recombinant Human CCK₂Cholecystokinin Receptor Membrane Preparation

半岛bd体育手机客户端 说明

一般描述

Full-length human CCKB cDNA encoding CCK₂
Cholecystokinins are a series of peptides of heterogeneous length (5 to 58 amino acids) that are derived from preprocholecystokinin and are found in gastrointestinal tissues and the central nervous system. Gastrin is a related peptide with 5 C-terminal amino acids identical to those of cholecystokinin. Two GPCRs, CCK₁(CCK_A) and CCK₂(CCK_B), bind to CCK and/or gastrin to mediate the biological effects of the peptides. CCK₂binds to CCK and gastrin with similar affinity, whereas CCK₁selectively binds sulfated CCK. Binding of ligands to CCK₂stimulates mobilization of intracellular calcium by activation of G_q/11. In the periphery, CCK₂is present in the stomach, where it mediates gastrin-stimulated gastric acid secretion. In the CNS, CCK₂has been implicated in anxiety, depression, schizophrenia, depression and opioid analgesia (Noble et al., 1999). Chemicon′s CCK₂membrane 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 CCK2 interactions and its ligands. The membrane preparations exhibit a Kd of 0.47 nM for [¹²⁵I]-cholecystokinin octapeptide (CCK-8). With 0.3 nM [¹²⁵I]CCK-8, 5 or 10 μg/well CCK₂Membrane Prep yields greater than 20-fold signal-to-background ratio.

应用

Radioligand binding assay and GTPγS binding.

生化/生理作用

Protein Target: CCK2 / CCKb
GPCR Class: A
Target Sub-Family: Cholecystokinin

质量

Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CCK₂membrane prep:

	10 ?g/well	5 ?g/well
Signal:Background	27.4	27.8
Specific Binding (cpm)	25,302	20,111

Specifications:
1unit = 5 μg
B _maxfor [ ¹²⁵I] CCK-8 binding: 3.47 pmol/mg protein
K _dfor [ ¹²⁵I] CCK-8 binding: ~0.47 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C

? Radioligand: [¹²⁵I]-CCK-8 (Perkin Elmer#:NEX-203 )

? 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 a unit is the amount of membrane that will yield greater than 20-fold signal:background with¹²⁵I labeled CCK-8 at 0.3 nM

外形

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with 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-1 cells
manufacturer/tradename	ChemiScreen Chemicon?
technique(s)	ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable
NCBI登记号	NM_176875.2
UniProt登记号	P32239
运输	dry ice
Gene Information	human ... CCKBR(887)

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