"ADORA1" Related Products

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Human ADORA1/Galpha15 Stable Cell Line-Chem-3

Cat.No.: CSC-RG0600
Cell Line Description: This 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 Galpha15 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.
Background: 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. A1 is also expressed in kidney, where it contributes to tubuloglomerular feedback.
Growth Properties: Adherent
Host Cell: Chem-3
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific silencing of ADORA1.2. Protein expression: ADORA1 in this cell line has been validated by immunocytochemical staining.3. EC50 for calcium mobilization by CPA: ~ 0.8 nM; IC50 for DPCPX: ~250 nM; Signal:noise ratio at CPA Emax: 747.
Application: Calcium flux assay, ligand binding assays
Sub-type: Adenosine
Propagation: Complete growth medium: RPMI-1640 containing 0.3 g/L L-glutamine, no pyruvate + 10% heat-inactivated FBS + 100 U/ml penicillin + 100 U/ml streptomycin + 1% Chem-3 Growth Supplement + G-418/Geneticin (600ug/mL)Note: The Chem-3 Growth Supplement is required for growth of Chem-3 cell linesPlating medium: RPMI-1640 containing 0.3 g/L L-glutamine, no pyruvate + 10% heat-inactivated FBS + 100 U/ml penicillin + 100 U/ml streptomycinAtmosphere: air, 95%; carbon dioxide (CO2), 5%. Temperature: 37 °C
Starting Cells From Frozen Cell Stock: 1. Immediately upon receipt, thaw cells or place cells in liquid nitrogen. Maintain frozen in liquid nitrogen for up to 5 years. 2. 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 conical tube containing 10 mL Growth Media. Centrifuge at 200 x g for 5 min. Remove the supernatant and resuspend the cell pellet in 20 mL Growth Media. Transfer to a T75 flask and place the flask in a humidified incubator at 37 degree °C with 5% CO2.
Subculturing: 1. Maintain the cells in suspension between 0.1 x 10^6 and 1 x 10^6 cells/mL. 2. Cells are typically passaged 1:10 every 2-3 days, or 1:20 every 3-4 days. Passaging ratio may be varied according to requirements of the investigator. Chem-3 cells tend to proliferate rapidly, so care should be taken not to allow them to become overconfluent.
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Freezing medium: RPMI-1640 containing 0.3 g/L L-glutamine, no pyruvate + 20% heat-inactivated FBS + 10% DMSO
Storage: Liquid nitrogen
Preservation: 1. Frozen stocks of cells should be prepared at the earliest passage possible after thawing, as follows: Count cells. Centrifuge cells at 200 x g for 5 min. Resuspend cells at 2-5 x 10^6 cells/mL in Freezing Media. Dispense 1 mL aliquots into cryopreservation vials. Freeze the cells by a controlled rate process, such as in an isopropanol-jacketed container placed at –70°C overnight. Store the vials in liquid nitrogen. 2. Cells should be passaged at least once after thawing prior to use in calcium flux assays.
Safety Considerations: The following safety precautions should be observed.1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.2. No eating, drinking or smoking while handling the stable line.3. Wash hands after handling the stable line and before leaving the lab.4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.5. All waste should be considered hazardous.6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship: Dry ice
Gene Name: ADORA1 adenosine A1 receptor [ Homo sapiens ]
Official Symbol: ADORA1
Synonyms: ADORA1; adenosine A1 receptor; adenosine receptor A1; RDC7;
Gene ID: 134
mRNA Refseq: NM_000674
Protein Refseq: NP_000665
MIM: 102775
UniProt ID: P30542
Chromosome Location: 1q32.1
Pathway: Adenosine P1 receptors, organism-specific biosystem; Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; G alpha (i) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; GPCR ligand binding, organism-specific biosystem; GPCRs, Class A Rhodopsin-like, organism-specific biosystem; Morphine addiction, organism-specific biosystem;
Function: G-protein beta/gamma-subunit complex binding; G-protein coupled adenosine receptor activity; G-protein coupled receptor binding; heterotrimeric G-protein binding; phospholipase C activity; protein binding; protein heterodimerization activity; purine nucleoside binding; receptor activity; signal transducer activity;

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Optional requirements on this protein    +Expand
C-fusion    N-fusion   Non-tagged
His    GST   Fc   Others
<1.0 eu/μg    <0.1 eu/μg   <0.01 eu/μg   Not required
Monomer Isolation    Dimer Isolation    Not required
>80% by SDS-PAGE    >90% by SDS-PAGE   >95% by SDS-PAGE   Others

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