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Calcium flux in D1-expressing Chem-6 cell line induced by A68930. D1-expressing Chem-6 cells and Wild-Type Chem-6 cells were lo...
ChemiScreen™ Human D1 Dopamine Receptor Calcium-Optimized FLIPR Cell Line
Description:
ChemiScreen™ Human D1 Dopamine Receptor Calcium-Optimized FLIPR Cell Line
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Qty/Pk:
2 vials
Product Overview:
Full-length human DRD1 cDNA encoding D1
Background Information:
Dopamine exerts its action by binding to five distinct dopamine receptors, all of which belong to G protein-coupled receptor family (Missale et al. 1998). The D1 subtype is the most abundant dopamine receptor in the central nervous system. Activation of D1 receptor stimulates adenylyl cyclase, activates cyclic AMP-dependent protein kinases. It regulates neuronal growth and development, mediate some behavioral responses and modulate dopamine receptor D2-mediated events. Chemicon's cloned human D1-expressing cell line is made in the Chem-6 host, which supports high levels of recombinant D1 expression on the cell surface and contains high levels of the promiscuous G protein to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between D1 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 the D1 subtype of the dopamine receptor. The D1 subtype is the most abundant dopamine receptor in the central nervous system. This G-protein coupled receptor stimulates adenylyl cyclase and activates cyclic AMP-dependent protein kinases. D1 receptors regulate neuronal growth and development, mediate some behavioral responses, and modulate dopamine receptor D2-mediated events. Alternate transcription initiation sites result in two transcript variants of this gene.
UniProt Summary:
FUNCTION: SwissProt: P21728 # This is one of the five types (D1 to D5) of receptors for dopamine. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase.
SIZE: 446 amino acids; 49293 Da
SUBUNIT: Interacts with DNAJC14 via its C-terminus (By similarity). Interacts with DRD1IP.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein (By similarity). Endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). Note=Transport from the endoplasmic reticulum to the cell surface is regulated by interaction with DNAJC14 (By similarity).
TISSUE SPECIFICITY: Detected in caudate, nucleus accumbens and in the olfactory tubercle.
SIMILARITY: SwissProt: P21728 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 446 amino acids; 49293 Da
SUBUNIT: Interacts with DNAJC14 via its C-terminus (By similarity). Interacts with DRD1IP.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein (By similarity). Endoplasmic reticulum membrane; Multi-pass membrane protein (By similarity). Note=Transport from the endoplasmic reticulum to the cell surface is regulated by interaction with DNAJC14 (By similarity).
TISSUE SPECIFICITY: Detected in caudate, nucleus accumbens and in the olfactory tubercle.
SIMILARITY: SwissProt: P21728 ## Belongs to the G-protein coupled receptor 1 family.
Species:
Human
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by A68930: ~ 0.097 nM
Signal:noise at agonist Emax: 452
Signal:noise at agonist Emax: 452
Cell Type:
Chem-6
Cell Line Type:
GPCR Cell Lines
Brand Family:
Chemicon
Host Cells:
Chem-6
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:
D1
UniProt Number:
Ligand Type:
non-peptide
Target Sub-Family:
Dopamine
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, 2 x 106 cells/mL
Gene Symbol:
- DRD1
- D
- DRD1A
- DADR
Protein or Enzyme Type:
GPCRs
Product Name:
ChemiScreen™ Human D1 Dopamine Receptor Calcium-Optimized FLIPR Cell Line
Entrez Gene Number:
Materials Required but Not Delivered:
F-12K Nutrient Mixture, Kaighn's Modification /10% heat inactivated fetal bovine serum/0.25 mg/mL Geneticin (G418)/0.2mg/mL Zeocin/100 U/mL each penicillin and streptomycin.