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ChemiScreen™ AT1 Calcium-Optimized FLIPR Cell Lines
Description:
ChemiScreen™ AT1 Calcium-Optimized FLIPR Cell Lines
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Qty/Pk:
2 vials
Product Overview:
Full-length human AGTR1 cDNA, encoding AT1
Background Information:
Angiotensin II (Ang II), an octapeptide produced by cleavage of angiotensinogen by angiotensin-converting enzyme, plays a fundamental role in cardiovascular homeostasis. Two GPCRs, AT1 and AT2, mediate the effects of AngII, although AT1 is primarily responsible for the effects of Ang II on renal function and development, thirst, electrolyte and water balance, and arterial blood pressure. Binding of Ang II to AT1 activates both Gq and Gi (De Gasparo et al., 2000). Chemicon's cloned human AT1-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant AT1 expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to enhance coupling of the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between AT1 and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays.
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:
Angiotensin II is a potent vasopressor hormone and a primary regulator of aldosterone secretion. It is an important effector controlling blood pressure and volume in the cardiovascular system. It acts through at least two types of receptors. This gene encodes the type 1 receptor which is thought to mediate the major cardiovascular effects of angiotensin II. This gene may play role in the generation of reperfusion arrhythmias following restoration of blood flow to ischemic or infarcted myocardium. It was previously thought that a related gene, denoted as AGTR1B, existed; however, it is now believed that there is only one type 1 receptor gene in humans. At least four transcript variants have been described for this gene. Additional variants have been described but their full-length nature has not been determined. The entire coding sequence is contained in the terminal exon and is present in all transcript variants.
UniProt Summary:
FUNCTION: SwissProt: P30556 # Receptor for angiotensin II. Mediates its action by association with G proteins that activate a phosphatidylinositol- calcium second messenger system.
SIZE: 359 amino acids; 41061 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Liver, lung, adrenal and adrenocortical adenomas.
PTM: C-terminal Ser or Thr residues may be phosphorylated.
DISEASE: SwissProt: P30556 # Defects in AGTR1 are a cause of renal tubular dysgenesis (RTD) [MIM:267430]. RTD is an autosomal recessive severe disorder of renal tubular development characterized by persistent fetal anuria and perinatal death, probably due to pulmonary hypoplasia from early-onset oligohydramnios (the Potter phenotype).
SIMILARITY: SwissProt: P30556 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 359 amino acids; 41061 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Liver, lung, adrenal and adrenocortical adenomas.
PTM: C-terminal Ser or Thr residues may be phosphorylated.
DISEASE: SwissProt: P30556 # Defects in AGTR1 are a cause of renal tubular dysgenesis (RTD) [MIM:267430]. RTD is an autosomal recessive severe disorder of renal tubular development characterized by persistent fetal anuria and perinatal death, probably due to pulmonary hypoplasia from early-onset oligohydramnios (the Potter phenotype).
SIMILARITY: SwissProt: P30556 ## Belongs to the G-protein coupled receptor 1 family.
Species:
Human
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by Angiotensin II: ~ 8.6 nM
Cell Type:
Chem-1
Cell Line Type:
GPCR Cell Lines
Brand Family:
Chemicon
Host Cells:
Chem-1
Presentation:
Cells are frozen at 2 x 10e6 cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.
Protein Target:
AT1
UniProt Number:
Ligand Type:
peptide
Target Sub-Family:
Angiotensin
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 37ºC 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 Ca++ and Mg++-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:
- AGTR1
- AT2R1A
- AT2R1B
- AGTR1B
- AT2R1
- AT1BR
- AGTR1A
- AT1AR
- AG2S
- HAT1R
- AT1
- AT1B
Protein or Enzyme Type:
GPCRs
Product Name:
ChemiScreen™ AT1 Calcium-Optimized FLIPR Cell Lines
Entrez Gene Number:
Materials Required but Not Delivered:
GROWTH MEDIA: 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 10e6 cells/mL