"Ffar1" Related Products

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Rat Ffar1-FLAG Stable Cell Line-HEK293T

Cat.No.: CSC-RG0251
Cell Line Description: HEK293T-RatFfar1-FLAG cell line is a hypotriploid human cell line, which has been transfected with a rat G-protein coupled receptor 40 (Ffar1) tagged in the N-terminus with FLAG to allow stably express of the rat Ffar1 tagged in the N-terminus with FLAG protein. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: G-protein coupled receptor 40 (GPR40 or FFA1) is specifically expressed in brain and pancreas. In pancreas, abundant GPR40 is localized to insulin-producing beta cells. Long-chain FFAs amplify glucose-stimulated insulin secretion from pancreatic beta cells by activating GPR40, indicating that GPR40 agonists and/or antagonists have potential for the development of new anti-diabetic drugs. GPR40 overexpression in breast cancer cells amplified oleate-induced proliferation, whereas silencing the GPR40 gene decreased it. These results suggest that GPR40 is implicated in the control of breast cancer cell growth by fatty acids and that GPR40 may provide a link between fat and cancer.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293T
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of rat GPR40.2. Protein expression: GPR40 expression in this cell line has been validated by WB.3. Functional validation: Functional assays.
Sub-type: Free Fatty Acid
Propagation: Complete growth medium: DMEM, 10% FBS, 1 μg/mL puromycinAtmosphere: 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 1:10 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: Ffar1 free fatty acid receptor 1 [ Rattus norvegicus ]
Official Symbol: Ffar1
Synonyms: FFAR1; free fatty acid receptor 1; G protein-coupled receptor 40; G-protein coupled receptor 40; Gpr40;
Gene ID: 266607
mRNA Refseq: NM_153304
Protein Refseq: NP_695216
Pathway: G alpha (q) signalling events, organism-specific biosystem; GPCR downstream signaling, organism-specific biosystem; GPCRs, Class A Rhodopsin-like, organism-specific biosystem; Incretin Synthesis, Secretion, and Inactivation, organism-specific biosystem; Integration of energy metabolism, organism-specific biosystem; Metabolism, organism-specific biosystem; Regulation of Insulin Secretion, organism-specific biosystem;
Function: G-protein coupled receptor activity; fatty acid binding; receptor activity; signal transducer activity;

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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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