生物源
human
品質等級
重組細胞
expressed in Chem-1 cells
製造商／商標名
ChemiScreen
Chemicon®
技術
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
NCBI登錄號
UniProt登錄號
運輸包裝
dry ice
一般說明
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.
Human CCK1 Receptor
應用
Radioligand binding assay and GTPγS binding
生化／生理作用
GPCR Class: A
Protein Target: CCK1 / CCKa
Target Sub-Family: Cholecystokinin
品質
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CCK1 Receptor membrane prep.
SPECIFICATIONS: 1 unit = 10 µg membrane preparation
Bmax: 0.15 pmol/mg
Kd: 0.5 nM
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 5.31 | 6.16 |
| Specific Binding (cpm) | 1274.9 | 1691.4 |
Bmax: 0.15 pmol/mg
Kd: 0.5 nM
規格
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.
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.
外觀
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.
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.
儲存和穩定性
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
法律資訊
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
免責聲明
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.
儲存類別代碼
12 - Non Combustible Liquids
水污染物質分類（WGK）
WGK 1
閃點（°F）
Not applicable
閃點（°C）
Not applicable
我们的科学家团队拥有各种研究领域经验，包括生命科学、材料科学、化学合成、色谱、分析及许多其他领域.
联系技术服务部门