"OPRK1" Related Products

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Human OPRK1-tGFP Stable Cell Line-U2OS

Cat.No.: CSC-RG0034
Cell Line Description: U2OS-HuOPRK1-tGFP cell line is a sarcoma cell line from human tibia, which has been transfected with a Human Kappa opioidReceptor (KOR) tagged in the N-terminus with tGFP to allow stably express of the human KOR tagged in the N-terminus with tGFP protein. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: kappa-opioid receptor is one of five related receptors that bind opium-like compounds in the brain and are responsible for mediating the effects of these compounds. These effects include altering the perception of pain, consciousness, motor control, and mood. Opioid receptors mediate the analgesic action and addictive properties of opioid drugs. The kappa-opioid receptor may provide a natural addiction control mechanism, and consequently selective agonists of this receptor may have therapeutic potential in the treatment of addiction.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: U2OS
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human OPRK1.2. Functional validation: Activation and Internalization assay for KOR-tGFP(Ec50 =3.42 x 10^-10M)
Sub-type: Opioid
Propagation: Complete growth medium: McCoy"s 5a medium with 1.5 mM Glutamine; 10% Foetal Bovine Serum (FBS).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 1:4 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: OPRK1 opioid receptor, kappa 1 [ Homo sapiens ]
Official Symbol: OPRK1
Synonyms: OPRK1; opioid receptor, kappa 1; kappa-type opioid receptor; KOR; KOR-1; K-OR-1; kappa opioid receptor; Opiate receptor, kappa-1; OPRK;
Gene ID: 4986
mRNA Refseq: NM_000912
Protein Refseq: NP_000903
MIM: 165196
UniProt ID: P41145
Chromosome Location: 8q11.2
Pathway: 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; Neuroactive ligand-receptor interaction, organism-specific biosystem; Neuroactive ligand-receptor interaction, conserved biosystem;
Function: G-protein coupled receptor activity; dynorphin receptor activity; protein binding; receptor activity; signal transducer activity;

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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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