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Figure 1. Calcium flux in A1-expressing Chem-3 cell line induced by N6-cyclopentyladenosine (CPA). A1-expressing Chem-3 cells ...
Figure 2. Assay for antagonist activity at A1 by calcium flux assay. A1-expressing Chem-3 cells were loaded with Fluo-4 and washed. DPCPX was added...
ChemiScreen™ A1 Calcium-Optimized FLIPR Cell Lines
Description:
ChemiScreen™ A1 Calcium-Optimized FLIPR Cell Lines
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Qty/Pk:
2 vials
Product Overview:
TRANSFECTION: Full-length human ADORA1 cDNA encoding A1
Background Information:
Extracellular adenosine mediates a multitude of biological effects, including wakefulness, antiarrythmia, bronchoconstriction and response to ischemia and oxidative stress. A family of four GPCR adenosine receptors, A1, A2A, A2B and A3, is responsible for these effects. The A1 receptor, which couples to Gi/o, is most highly expressed in brain, and mediates endogenous antinociception and neuronal response to hypoxia (Fredholm et al., 2001). A1 is also expressed in kidney, where it contributes to tubuloglomerular feedback. Chemicon's cloned human A1-expressing cell line is made in the Chem-3 host, which supports high levels of recombinant A1 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 A1 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:
The protein encoded by this gene is an adenosine receptor that belongs to the G-protein coupled receptor 1 family. There are 3 types of adenosine receptors, each with a specific pattern of ligand binding and tissue distribution, and together they regulate a diverse set of physiologic functions. The type A1 receptors inhibit adenylyl cyclase, and play a role in the fertilization process. Animal studies also suggest a role for A1 receptors in kidney function and ethanol intoxication. Transcript variants with alternative splicing in the 5' UTR have been found for this gene.
UniProt Summary:
FUNCTION: SwissProt: P30542 # Receptor for adenosine. The activity of this receptor is mediated by G proteins which inhibit adenylyl cyclase.
SIZE: 326 amino acids; 36512 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P30542 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 326 amino acids; 36512 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P30542 ## Belongs to the G-protein coupled receptor 1 family.
Species:
Human
Brand Family:
Chemicon
Protein Target:
A1
Presentation:
Cells are frozen at 2 x 106 cells/mL in RPMI/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.
Ligand Type:
non-peptide
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 sterile conical tube containing 10 mL growth media, and centrifuge at 200 x g. Remove supernatant and resuspend in fresh growth media. Transfer to a T75 flask containing 20 mL final volume growth media. Cells should be maintained between 0.1 x 106 and 1.0 x 106 cells/mL, and are typically passaged 1:10 every 3-4 days. Cells should be passaged at least once after thawing prior to use in calcium flux assays.
Target Sub-Family:
Adenosine
GPCR Class:
A
Promotional Text:
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Gene Symbol:
- ADORA1
- RDC7
Protein or Enzyme Type:
GPCRs
Product Name:
ChemiScreen™ A1 Calcium-Optimized FLIPR Cell Lines
Materials Required but Not Delivered:
GROWTH MEDIA: RPMI containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/0.8 mg/ml Geneticin (G418)/100 U/ml each penicillin and streptomycin, containing 1:100 dilution of Chem-3 Growth Supplement (Chemicon catalog # HTSCHEM-3S).
Concentration:
2 x 106 cells/mL
Quality Assurance:
SPECIFICATIONS:
EC50 for calcium mobilization by CPA: ~ 0.8 nM
IC50 for DPCPX: ~250 nM
Signal:noise ratio at CPA Emax: 747
EC50 for calcium mobilization by CPA: ~ 0.8 nM
IC50 for DPCPX: ~250 nM
Signal:noise ratio at CPA Emax: 747
Cell Type:
Chem-3
Cell Line Type:
GPCR Cell Lines
Host Cells:
Chem-3
Format:
Cell Line
UniProt Number:
Packaging:
1 mL per vial
Entrez Gene Number: