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Human DRD2-SNAP Stable Cell Line-CHO

Cat.No.: CSC-RG0047
Cell Line Description: CHO-HuDRD2-SNAP cell line is clonally-derived from a CHO cell line, which has been transfected with a Human dopamine receptor D2 (DRD2) tagged in the N-terminus with SNAP to allow stably express of the human DRD2 tagged in the N-terminus with SNAP protein. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: This gene encodes the D2 subtype of the dopamine receptor. This G-protein coupled receptor inhibits adenylyl cyclase activity. A missense mutation in this gene causes myoclonus dystonia; other mutations have been associated with schizophrenia. Alternative splicing of this gene results in two transcript variants encoding different isoforms. A third variant has been described, but it has not been determined whether this form is normal or due to aberrant splicing.
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human DRD2.2. Protein expression: DRD2 expression in this cell line has been validated by WB.3. Functional validation: Ligand binding assay; Receptor dimerization assays; cAMP assay; Internalization assay
Sub-type: Dopamine
Propagation: Complete growth medium: Ham"s F12, 10% dFBS, 50U/ml penicillin, 50μg/ml streptomycin,2mM Hepes Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock: Thaw the vials of cryopreserved cells rapidly in a water bath at 37°C. Carefully transfer the cells to a cell culture vessel containing the appropriate volume of pre-warmed culture medium without G418 so that cell density reaches 4 x 10^4 to 8 x 10^4 cells/cm2. The day after, ensure that cells have properly attached and carefully replace with fresh culture medium supplemented with G418. Allow cells to grow until 80% confluency then transfer in a new cell culture vessel. Note that final DMSO concentration must not exceed 0.5%.
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.6. Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 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: DRD2 dopamine receptor D2 [ Homo sapiens ]
Official Symbol: DRD2
Synonyms: DRD2; dopamine receptor D2; D dopamine receptor; dopamine D2 receptor; dopamine receptor D2 isoform; seven transmembrane helix receptor; D2R; D2DR;
Gene ID: 1813
mRNA Refseq: NM_000795
Protein Refseq: NP_000786
MIM: 126450
UniProt ID: P14416
Chromosome Location: 11q22-q23
Pathway: Amine ligand-binding receptors, organism-specific biosystem; Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; Cocaine addiction, organism-specific biosystem; Cocaine addiction, conserved biosystem; Dopamine receptors, organism-specific biosystem; Dopaminergic synapse, organism-specific biosystem; Dopaminergic synapse, conserved biosystem;
Function: G-protein coupled receptor activity; dopamine binding; dopamine receptor activity, coupled via Gi/Go; dopamine receptor activity, coupled via Gi/Go; drug binding; drug binding; ionotropic glutamate receptor binding; potassium channel regulator activity; protein binding; receptor activity; receptor binding; signal transducer activity;

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