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ChemiSCREEN Membrane Preparation Recombinant Human CCK1 Cholecystokinin Receptor
产品编号: | 4094858 |
规格: | Human CCK1 / CCKa GPCR membrane preparation for Radioligand binding Assays & GTPγS binding. |
包装规格: | 200 UNITS |
产品类别: | 进口试剂 |
品牌: | Sigma-Aldrich |
优惠价: | 立即咨询 |
基本信息
eCl@ss | 32161000 | |||||||||
NACRES | NA.41 | |||||||||
General description【一般描述】 | Human CCK1 Receptor 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 (CCKA) and CCK2 (CCKB), bind to CCK and/or gastrin to mediate the biological effects of the peptides. CCK1 selectively binds sulfated CCK, whereas CCK2 binds to CCK and gastrin with similar affinity. Binding of ligands to CCK1 stimulates mobilization of intracellular calcium by activation of Gq/11. CCK1 receptors in the periphery are primarily localized in the pancreas, gallbladder, pylorus, intestine where they are responsible for the regulation of diverse digestive processes. They are also present in select areas of the peripheral nervous system (vagus nerve), and the CNS where they mediate the satiety effects of CCK, regulate an increase in dopamine release, and antagonize opioid analgesia (Noble et al., 1999). Chemicon′s CCK1 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 CCK1 interactions and its ligands. The membrane preparations exhibit a Kd of 0.5 nM for [125I]-CCK-8 sulfated. With 10.0ug/well CCK1 membrane prep and 0.5 nM [125I]-CCK-8 sulfated, a greater than 4-fold signal-to-background ratio was obtained. | |||||||||
Application【应用】 | Radioligand binding assay and GTPγS binding | |||||||||
Biochem/physiol Actions【生化/生理作用】 | Protein Target: CCK1 / CCKa GPCR Class: A Target Sub-Family: Cholecystokinin | |||||||||
Quality【质量】 | Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CCK1 Receptor membrane prep.
Bmax: 0.15 pmol/mg Kd: 0.5 nM | |||||||||
Specifications【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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is blocked 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. Note: Due to the acidic property of the labeled sulfated CCK-8, filter plates coated with PEI bind the labeled ligand nonspecifically and result in elevated backgrounds. 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] -CCK-8 sulfated (Perkin Elmer # NEX203) Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, 0.1% BSA, filtered and stored at 4°C. | |||||||||
Physical form【外形】 | One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 4-fold signal:background with 125I-labeled CCK-8 at 0.5 nM. Liquid in packaging buffer: 50 mM Tris, pH 7.4, 10% glycerol with no preservatives. Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC. | |||||||||
Legal Information【法律信息】 | CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany |
产品性质
Quality Level【质量水平】 | 100 |
biological source【生物来源】 | human |
recombinant【重组】 | expressed in Chem-1 cells |
manufacturer/tradename | ChemiScreen Chemicon® |
technique(s) | ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable |
NCBI accession no.【NCBI登记号】 | NM_000730.2 |
UniProt accession no.【UniProt登记号】 | P32238 |
shipped in【运输】 | dry ice |
产品说明
Storage and Stability【储存及稳定性】 | Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw. |
Disclaimer【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. |
安全信息
Storage Class Code【储存分类代码】 | 12 - Non Combustible Liquids |
WGK | WGK 1 |