ChemiSCREEN Human VPAC₂VIP Receptor Membrane Preparation

半岛bd体育手机客户端 说明

一般描述

Vasoactive intestinal peptide (VIP), a 28 amino acid peptide originally isolated by its vasodilation activity, binds to two class B GPCRs, VPAC₁and VPAC₂, to exert its functions in the CNS, vasculature, immune system and adrenal medulla (Harmar et al., 1998). In the immune system, VPAC₂is expressed on stimulated CD4 T cells, and binding of T cell-derived VIP to VPAC₂induces a shift toward the Th2 pathway (Delgado et al., 2004; Voice et al., 2004). In addition, VPAC2 is an essential regulator of the circadian pacemaker of the hypothalamic suprachiasmatic nuclei (Hughes et al., 2004). Chemicon′s VPAC₂membrane 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 VPAC₂interactions with PACAP. The membrane preparations exhibit a Kd of 0.9 nM for [¹²⁵I]-PACAP27. With 5 ?g/well VPAC₂Membrane Prep and 0.5 nM [¹²⁵I]-PACAP27, a greater than 15-fold signal-to-background ratio was obtained.
Human VPAC2

应用

Radioligand binding assay, and GTPγS binding.

生化/生理作用

Target Sub-Family: VIP/PACAP
GPCR Class: B
Protein Target: VPAC2 / VIP2

质量

Signal:background and specific binding values obtained in a competition binding assay with 5ug/well of VPAC2 membrane prep:

	10 ?g/well	5 ?g/well
Signal:Background	25.5.	22.8
Specific Binding (cpm)	32913	27650

SPECIFICATIONS: 1 unit = 5 ?g membrane preparation
Bmax: 17.3 pmol/mg
Kd: 0.9 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] PACAP27 (Perkin Elmer # NEX294)
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 an unit is the amount of membrane that will yield greater than 15-fold signal:background with¹²⁵I-labeled PACAP27 at 0.5 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 the indicated concentration in 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
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_003382.3
UniProt登记号	P41587
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