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Figure 2. Inhibition of Gastrin I-induced calcium mobilization in CCK2-expressing Chem-1 cells by YM 022. Chem-1 cells were loaded with Fluo-4 and p...
Figure 1. Calcium flux in CCK2-expressing Chem-1 cell line induced by Gastrin I. CCK2-expressing Chem-1 cells and Wild-Type Chem-1 cells (Chemicon c...
ChemiScreen™ CCK2 Cholecystokinin Receptor Calcium-Optimized FLIPR Cell Lines
Description:
ChemiScreen™ CCK2 Cholecystokinin Receptor Calcium-Optimized FLIPR Cell Lines
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Qty/Pk:
2 vials
Product Overview:
Full-length human CCKB cDNA encoding CCK2
Background Information:
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. CCK2 binds to CCK and gastrin with similar affinity, whereas CCK1 selectively binds sulfated CCK. Binding of ligands to CCK2 stimulates mobilization of intracellular calcium by activation of Gq/11. In the periphery, CCK2 is present in the stomach, where it mediates gastrin-stimulated gastric acid secretion. In the CNS, CCK2 has been implicated in anxiety, depression, schizophrenia, depression and opioid analgesia (Noble et al., 1999). Chemicon's cloned human CCK2-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant CCK2 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 antagonists of interactions between the CCK2 and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays
Species Reactivity:
Human
Usage Statement:
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.
Entrez Gene Summary:
This gene encodes a G-protein coupled receptor for gastrin and cholecystokinin (CCK), regulatory peptides of the brain and gastrointestinal tract. This protein is a type B gastrin receptor, which has a high affinity for both sulfated and nonsulfated CCK analogs and is found principally in the central nervous system and the gastrointestinal tract. A misspliced transcript variant including an intron has been observed in cells from colorectal and pancreatic tumors.
UniProt Summary:
FUNCTION: SwissProt: P32239 # Receptor for gastrin and cholecystokinin. The CKK-B receptors occur throughout the central nervous system where they modulate anxiety, analgesia, arousal, and neuroleptic activity. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system.
SIZE: 447 amino acids; 48419 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P32239 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 447 amino acids; 48419 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P32239 ## Belongs to the G-protein coupled receptor 1 family.
Species:
Human
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by Gastrin I: ~ 3.1 nM
IC50 for YM 022: 22 nM
Signal:noise ratio at Gastrin I Emax: 623
IC50 for YM 022: 22 nM
Signal:noise ratio at Gastrin I Emax: 623
Cell Type:
Chem-1
Cell Line Type:
GPCR Cell Lines
Brand Family:
Chemicon
Host Cells:
Chem-1
Presentation:
Cells are frozen at 2 x 106 cells/mL in DMEM/20% fetal bovine serum/100 U/mL penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.
Protein Target:
CCK2 / CCKb
UniProt Number:
Ligand Type:
peptide
Target Sub-Family:
Cholecystokinin
Storage Conditions:
Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37°C water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO2. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca2+ and Mg2+-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.
Promotional Text:
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GPCR Class:
A
Packaging:
1 mL per vial
Gene Symbol:
- CCKBR
- CCK-BR
- CCK2R
- CCK-B
- GASR
- CCKRB
- CCK2-R
Protein or Enzyme Type:
GPCRs
Product Name:
ChemiScreen™ CCK2 Cholecystokinin Receptor Calcium-Optimized FLIPR Cell Lines
Entrez Gene Number:
Materials Required but Not Delivered:
DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/mL Geneticin (G418)/100 U/mL each penicillin and streptomycin
Concentration:
2 x 106 cells/mL