merck millipore,默克密理博,HTS052M,ChemiScreen™ A3 Membrane Preparation

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merck millipore,默克密理博,HTS052M,ChemiScreen™ A3 Membrane Preparation
产品名称：ChemiScreen™ A3 Membrane Preparation
产品型号：HTS052M
Human A3 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

merck millipore,默克密理博,HTS052M,ChemiScreen™ A3 Membrane Preparation

产品介绍

merck millipore,默克密理博,HTS052M,ChemiScreen™ A3 Membrane Preparation

重要规格表

应用说明	品种	宿主细胞	蛋白靶
Radioligand binding assay and GTPγS binding.	Human	Chem-3	A3

描述
产品目录编号	HTS052M
品牌系列	Chemicon®
商名	Chemicon ChemiScreen
描述	ChemiScreen™ A3 Membrane Preparation
概述	Human A3
背景信息	Extracellular adenosine mediates a multitude of biological effects, including wakefulness, antiarrythmia, bronchoconstriction and response to ischemia and oxidative stress. A family of four GPCR adenosine receptors, A1, A2A, A2B and A3, is responsible for these effects (Fredholm et al., 2001). A3, which couples to Gi/o, is expressed in mast cells along with A2B. Mice lacking A3 display reduced mast cell degranulation and bronchoconstriction in response to adenosine (Tilley et al., 2003; Zhong et al., 2003). Chemicon's cloned human A3-expressing cell line is made in the Chem-3 host, which supports high levels of recombinant A3 expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists at A3. Chemicon's A3 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 A3 interactions with its ligands. The membrane preparations exhibit a Kd of 4.7 nM for [¹²⁵I]-AB-MECA. With 5 mg/well A3 Membrane Prep and 5nM [¹²⁵I]-AB-MECA, a greater than 3-fold signal-to-background ratio is obtained.

产品信息
演示	Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.

应用
应用	Human A3 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
应用说明	Radioligand binding assay and GTPγS binding.

生物信息
浓缩	1 mL, 1.0 mg/mL
品种	Human
品种反应性	Human
Entrez基因编号	NM_020683.6 NM_000677.3 NM_001081976.1
Entrez基因汇总	This gene encodes a protein that belongs to the family of adenosine receptors, which are G-protein-coupled receptors that are involved in a variety of intracellular signaling pathways and physiological functions. The receptor encoded by this gene mediates a sustained cardioprotective function during cardiac ischemia, it is involved in the inhibition of neutrophil degranulation in neutrophil-mediated tissue injury, it has been implicated in both neuroprotective and neurodegenerative effects, and it may also mediate both cell proliferation and cell death. Multiple transcript variants encoding different isoforms have been found for this gene.
基因符号	A3AR ADORA3 AD026 bA552M11.5 ORFNames=UNQ1931/PRO4406 RP11-552M11.7
GPCR类别	A
宿主细胞	Chem-3
蛋白靶	A3
靶子系列	Adenosine
UniProt编号	P33765
UniProt汇总	FUNCTION: SwissProt: P33765 # Receptor for adenosine. The activity of this receptor is mediated by G proteins which inhibits adenylyl cyclase. Possible role in reproduction. SIZE: 318 amino acids; 36185 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. PTM: Phosphorylation on Ser-317 may be crucial for rapid desensitization (By similarity). SIMILARITY: SwissProt: P33765 ## Belongs to the G-protein coupled receptor 1 family.
孵化条件	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, and then washed with 50 mM Tris, pH 7.4. 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]-AB-MECA (Perkin Elmer# :NEX312) 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 3-fold signal:background with ¹²⁵I-labeled AB-MECA at 0.2 nM

产品使用声明

质量保证

Table 1. Signal:background and specific binding values obtained in a competition binding assay with A₃ membrane preparation:

	10 µg/well	5 µg/well
Signal:Background	3.7	4.0
Specific Binding (cpm)	1740	1708

SPECIFICATIONS:
1 unit = 5 μg
Bmax: 2.03 pmol/mg
Kd: 4.7 nM

Table 2. Summary of adenosine receptor agonists in a competition binding assay with A₃ membrane preparation:

agonist	Known selectivity	Observed IC50 (nM)
CI-IB-MECA	A3	1.3
NECA	Non-selective	135
CPA	A1	345
CGS21680	A2A	396

Table 3. Summary of adenosine receptor antagonists in a competition binding assay with A₃ membrane preparation:

antagonist	Known selectivity	Observed IC50 (nM)
VUF5574	A3	2.1
DPCPX	A1	759
MRS1754	A2B	1919
ZM241385	A2A	2513

使用声明

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.