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Home > Products > Transfected Stable Cell Lines > Protease-Activated > Human F2RL1 Stable Cell Line-CHO

Human F2RL1 Stable Cell Line-CHO

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His(3)
Cat.No.:
CSC-RG0921
Cell Line Description:
CHO-HuF2RL1 cell line is clonally-derived from a CHO cell line, which has been transfected with a human coagulation factor II (thrombin) receptor-like 1 F2RL1 to allow expression of the human F2RL1. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background:
Protease-activated receptor-2 (PAR2) belongs to a four-member family of GPCRs that are activated by proteases (see Ossovskaya et al for review). These proteases cleave a specific extracellular amino-terminal domain of the receptor to reveal a tethered ligand, which in turn activates the receptor which couples via Gq to release intracellular calcium (Nystedt, S. et al., 1995). PAR2 is expressed in the vasculature, GI tract, CNS, lung, kidney, liver, and heart. PAR2 is a relevant target for arthritis, inflammation, bowel disease, and cancer.
Growth Properties:
Adherent
Morphology:
Epithelial-like
Host Cell:
CHO-K1
Cell Line Validation:
1. Gene expression: qPCR experiments determined specific expression of human F2RL1.2. Protein expression: F2RL1 expression in this cell line has been validated by WB.3. Functional validation: binding, calcium assay.
Sub-type:
Protease-activated
Propagation:
Complete growth medium: Ham"s F12, 10% FBS, 1% NEAA, 0.4 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 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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