"PDE2A" Related Products

Human PDE2A Stable Cell Line-HEK293-CNG-Gs

Cat.No.: CSC-RG0923
Cell Line Description: HEK293-CNG-Gs-HuPDE2A cell line is a hypotriploid human cell line, which has been transfected with a Human phosphodiesterase 2A, cGMP-stimulated (PDE2A) to allow stably express of the human PDE2A. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: These cells express a modified CNG (Cyclic Nucleotide Gated) channel that opens in response to elevated intracellular cAMP levels and consequently depolarizes cells. Cell depolarization can be easily measured with fluorescent Membrane Potential Sensive Dye using a plate reader or FLIPR and is directly related to intracellular cAMP content.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293-CNG-Gs
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific expression of human PDE2A.2. Protein expression: PDE2A expression in this cell line has been validated by WB.
Stability: The receptor specific activity is stable for 10 weeks continuous passage
Sub-type: Phosphodiesterase
Propagation: Complete growth medium: DMEM-10%FBS supplemented with 250 mg/ml G418, 1 mg/ml Puromycin and 5 mg/ml blasticidin.Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock: 1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37°C water bath for 2 minutes until nearly (80%) thawed. Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.2. Clean the outside of the vial with 70% ethanol before opening.3. Remove the cells from the vial and add slowly into a 15ml conical tube containing 10 ml pre-warmed media. 4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.6. Place the cells in the 37°C incubator with 5% CO2.7. Allow incubation for 3-4 days to reach confluence. The cells will re-attach to the surface over a period of several days in culture at 37°C.
Subculturing: 1. Remove and discard culture medium.2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.5. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended Medium Renewal: 2 to 3 times per week
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 90%; DMSO, 10%
Storage: Liquid nitrogen
Preservation: 1. Detach cells from culture dish according to the Sub-Culture Procedure.2. Resuspend cells at a density of 5 x 10^6 cells/mL in freeze medium.Note: A T-75 culture flask typically yields enough cells for preparing two frozen vials.3. Aliquot 1 mL cells into cryogenic vials.4. Place vials in a freezing container and store at –80 °C overnight.5. Transfer vials to liquid nitrogen for long term storage. If properly stored, cells should remain stable for years.
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: PDE2A phosphodiesterase 2A, cGMP-stimulated [ Homo sapiens ]
Official Symbol: PDE2A
Synonyms: PDE2A; phosphodiesterase 2A, cGMP-stimulated; cGMP-dependent 3,5-cyclic phosphodiesterase; cGMP-stimulated phosphodiesterase 1; cGMP-stimulated phosphodiesterase 4; cyclic GMP-stimulated phosphodiesterase; PDE2A1; PED2A4; cGSPDE; CGS-PDE;
Gene ID: 5138
mRNA Refseq: NM_001143839
Protein Refseq: NP_001137311
MIM: 602658
UniProt ID: O00408
Chromosome Location: 11q13.1-q14.1
Pathway: G alpha (s) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; Hemostasis, organism-specific biosystem; Morphine addiction, organism-specific biosystem; Morphine addiction, conserved biosystem; Nitric oxide stimulates guanylate cyclase, organism-specific biosystem; Platelet homeostasis, organism-specific biosystem;
Function: 3,5-cyclic-nucleotide phosphodiesterase activity; TPR domain binding; cAMP binding; cGMP binding; cGMP binding; cGMP-stimulated cyclic-nucleotide phosphodiesterase activity; calcium channel activity; cyclic-nucleotide phosphodiesterase activity; drug binding; drug binding; hydrolase activity; metal ion binding; nucleotide binding; phosphoric diester hydrolase activity; protein binding; protein homodimerization 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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