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Human ADRA2B Stable Cell Line-CHO

Cat.No.: CSC-RG0798
Cell Line Description: CHO-HuADRA2B cell line is clonally-derived from a CHO cell line, which has been transfected with a human adrenoceptor alpha 2B ADRA2B to allow expression of the human ADRA2B. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: Three alpha2-adrenoceptors have been cloned, and are designated alpha2A, alpha2B and alpha2C. All three receptors are activated by norepinephrine and all three are coupled via Gi to adenylate cyclase. The alpha2 adrenergic receptor is widely distributed in the CNS, lung, blood vessels and skeletal muscle and mediates vasoconstriction and modulation of neurotransmission.
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO-K1
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific expression of human ADRA2B.2. Protein expression: ADRA2B expression in this cell line has been validated by WB.3. Functional validation: binding, cAMP assay.
Stability: 12 passages of continuous culture (6 weeks at 2 passages/week)
Sub-type: Adrenoceptors
Propagation: Complete growth medium: Ham"s F12, 10% dFBS, 1x NEAA, 0.5 mg/mL G418Atmosphere: 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 1:4 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 95%; DMSO, 5%
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: ADRA2B adrenergic, alpha-2B-, receptor [ Homo sapiens ]
Official Symbol: ADRA2B
Synonyms: ADRA2B; adrenergic, alpha-2B-, receptor; ADRA2L1, ADRA2RL1; alpha-2B adrenergic receptor; ADRARL1; alpha-2BAR; alpha-2B adrenoceptor; alpha-2B adrenoreceptor; G-protein coupled receptor; adrenergic receptor alpha 2B; alpha-2B-adrenergic receptor; alpha-2-adrenergic receptor-like 1; ADRA2B adrenergic, alpha-2B-, receptor; alpha-2 adrenergic receptor subtype C2; ADRA2L1; ADRA2RL1; ALPHA2BAR;
Gene ID: 151
mRNA Refseq: NM_000682
Protein Refseq: NP_000673
MIM: 104260
UniProt ID: P18089
Chromosome Location: 2q11.1
Pathway: Adrenaline signalling through Alpha-2 adrenergic receptor, organism-specific biosystem; Adrenoceptors, organism-specific biosystem; Amine ligand-binding receptors, organism-specific biosystem; Class A/1 (Rhodopsin-like receptors), organism-specific biosystem; G alpha (i) signalling events, organism-specific biosystem; G alpha (z) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem;
Function: G-protein coupled receptor activity; adrenergic receptor activity; alpha2-adrenergic receptor activity; epinephrine binding; protein binding; receptor activity; signal transducer activity;

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