"SUCNR1" Related Products


Human SUCNR1-FLAG Stable Cell Line-CHO

Cat.No.: CSC-RG0233
Cell Line Description: CHO-HuSUCNR1-FLAG is clonally-derived from a CHO-K1 cell line which has been transfected with a human succinate receptor 1 (SUCNR1) tagged in the N-terminus with FLAG to allow stably express of the human SUCNR1 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: GPR91, also known as SUCNR1, is a G Protein-Coupled Receptor with 339 amino acids. It has been characterized as a receptor for Succinate, a citric acid cycle intermediate. Succinate plays a key role in energy metabolism. Local interstitial accumulation of Succinate has recently been reported to serve as an indicator of ischemic or diabetic organ damage in the brain, liver, and kidney. In diabetes patients, the accumulation of Succinate is detectable in the plasma, and more significantly in the renal tubular fluid and urine. It is therefore considered a potential new biomarker of local tissue damage. It has also been shown that Succinate increases blood pressure in animals. The Succinate-induced hypertensive effect involves the renin-angiotensin system that is shown to be absent in GPR91-deficient mice. There is a possible role for GPR91 in renovascular hypertension, a disease closely linked to atherosclerosis, diabetes and renal failure. In a recombinant system overexpressing GPR91, Succinate was shown to not only stimulate calcium mobilization and inositol phosphate (IP) accumulation through the stimulation of Galphaq pathway but also to activate the Erk1/2 MAPK pathway and inhibit forskolin-stimulated cAMP accumulation through Galphai pathway.
Growth Properties: Adherent
Morphology: Epithelial-like
Host Cell: CHO-K1
Cell Line Validation: 1. Gene expression: RT-PCR experiments determined specific expression of human SUCNR1.2. Protein expression: SUCNR1 expression in this cell line has been validated by WB.3. Functional validation: Functional assays.
Sub-type: GPR
Propagation: Complete growth medium: DMEM/F12, 10% FBS, 10 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 1:8 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: SUCNR1 succinate receptor 1 [ Homo sapiens ]
Official Symbol: SUCNR1
Synonyms: SUCNR1; succinate receptor 1; G protein coupled receptor 91 , GPR91; P2Y purinoceptor 1; G protein-coupled receptor 91; G-protein coupled receptor 91; GPR91;
Gene ID: 56670
mRNA Refseq: NM_033050
Protein Refseq: NP_149039
MIM: 606381
UniProt ID: Q9BXA5
Chromosome Location: 3q25.1
Pathway: GPCRs, Class A Rhodopsin-like, organism-specific biosystem;
Function: G-protein coupled purinergic nucleotide receptor activity; G-protein coupled receptor activity; receptor activity; signal transducer activity;

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