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Home > Products > Transfected Stable Cell Lines > SPC/LPC /Proton-Sensor > Human GPR84-FLAG Stable Cell Line-CHO-K1 Galphaqi5

Human GPR84-FLAG Stable Cell Line-CHO-K1 Galphaqi5

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Cat.No.:
CSC-RG0259
Cell Line Description:
CHO-K1 Galphaqi5-HuGPR84-FLAG is clonally-derived from a CHO-K1 cell line which has been transfected with a human G protein-coupled receptor 84 (GPR84) tagged in the N-terminus with FLAG to allow stably express of the human GPR84 tagged in the N-terminus with FLAG. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background:
Human GPR84 is a G protein-coupled receptor with 396 amino acids. The hgpr84 transcript was found in brain, heart, muscle, colon, thymus, spleen, kidney, liver, intestine, placenta, lung and leukocytes. A study in GPR84-deficient mice revealed a novel role for GPR84 in regulating early IL-4 gene expression in activated T cells in response to CD3 crosslinking.
Growth Properties:
Adherent
Morphology:
Epithelial-like
Host Cell:
CHO-K1 Galphaqi5
Cell Line Validation:
1. Gene expression: RT-PCR experiments determined specific expression of human GPR84.2. Protein expression: GPR84 expression in this cell line has been validated by WB.3. Functional validation: Functional assays.
Sub-type:
SPC/LPC /proton-sensor
Propagation:
Complete growth medium: DMEM/F12, 10% FBS, 10 μg/mL puromycin, 250μg/mL hygromycinAtmosphere: 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


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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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45-1 Ramsey Road, Shirley, NY 11967, USA
USA: 1-631-559-9269  1-631-448-7888
Europe: 44-207-097-1828
FAX: 1-631-938-8127
EMAIL: info@creative-biomart.com

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