"GPR119" Related Products


Human GPR119/Clytin Stable Cell Line-HEK293

Cat.No.: CSC-RG0715
Cell Line Description: These cells express a novel variant of clytin, a calcium-activated photoprotein, to enable sensitive luminescent detection of ligand-induced calcium flux. The clytin contains a mutation that increases its affinity for calcium to a level that permits detection of cytosolic calcium in many cells with greater sensitivity than other photoproteins targeted to the mitochondria. Luminescent calcium assays offer several advantages over fluorescent calcium assays including increased sensitivity and lack of interference from fluorescent compounds. These human GPR119 receptor-expressing cells were made by stable transfection of HEK293 cells with clytin and the receptor and a promiscuous G protein to couple the receptor to the calcium signaling pathway. These stability-tested cells are ready for luminescent analysis of agonists, antagonists and modulators at the GPR119 receptor.
Background: GPR119 is a GPCR that is predominately expressed in pancreatic beta cells and enteroendocrine cells in the GI tract in humans. Phospholipids and fatty acid amines are possible endogenous ligands, and oleylethanolamide is one of the more potent ligands known. Agonists of GPR119 elevate cyclic AMP levels in cell models which can potentiate glucose-stimulated insulin secretion. In mice, it has been showed that a GPR119 agonist stimulated the release of both GLP-1 and GIP in wildtype, but not GPR119-deficient animals. GPR119 agonists also improve glycaemic control for mice in vivo, and GLP-1 administration reduces food intake and results in weight loss in humans. Taken together, these data suggest that GPR119 could be an important target for therapeutic management of diabetes and obesity in humans.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific silencing of GPR119.2. Protein expression: GPR119 in this cell line has been validated by immunocytochemical staining.
Application: Calcium Flux Assays; cAMP Assays
Sub-type: Orphan
Propagation: Complete growth medium: DMEM/F12 with 2.5 mM glutamine, 10% heat-inactivated FBS, 1x Nonessential amino acids, 1x Pen-Strep, 250μg/mL Genetecin/G-418Plating medium: DMEM/F12 with 2.5 mM glutamine, 10% heat-inactivated FBS, 1x Nonessential amino acids, 1x Pen-StrepAtmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37 °C
Starting Cells From Frozen Cell Stock: 1. Immediately upon receipt, thaw cells or place cells in liquid nitrogen. 2. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37°C water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing growth media. Place the flask in a humidified incubator at 37°C with 5% CO2.3. After 8-24 h, all live cells will be attached. Viability of the cells is expected to be 50-80%. At this time, replace media to remove residual DMSO, and return to incubator.
Subculturing: 1. When cells are approximately 80% confluent, passage the cells as follows: Remove media and wash once with HBSS without Ca2+ and Mg2+ (10 mL/T75). Add Accutase at 1 mL/T75 and keep at room temperature until cells begin to round up and detach (5-10 minutes). Gently rap the side of the flask to dislodge the cells. Neutralize Accutase by addition of 4 mL Growth Media per 1 mL Accutase. 2. Cells are typically passaged 1:10 every 3-4 days. Passaging ratio may be varied according to requirements of the investigator.
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Freezing medium: 90% heat-inactivated FBS + 10% DMSO
Storage: Liquid nitrogen
Preservation: 1. Frozen stocks of cells should be prepared at the earliest passage possible after thawing, as follows: Count detached cells (prepared as in Subculture-Step 1). Centrifuge cells at 200 x g for 5 min. Resuspend cells at 5 x 10^6 cells/mL in Freezing Media (cell densities of 2-10 x 10^6 are also acceptable if necessary). Dispense 1 mL aliquots into cryopreservation vials. Freeze the cells by a controlled rate process, such as in an isopropanol-jacketed container placed at –70°C overnight. Store the vials in liquid nitrogen. 2. Use of cells immediately after thawing is feasible for some cell lines and is being further validated. Some cell lines may need to be passaged at least once after thawing prior to use in calcium flux assays. Cells should be resuspended in Plating Media for plating for calcium assay.
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: GPR119 G protein-coupled receptor 119 [ Homo sapiens ]
Official Symbol: GPR119
Synonyms: GPR119; G protein-coupled receptor 119; glucose-dependent insulinotropic receptor; GPCR2; hGPCR2; G-protein coupled receptor 2; G-protein coupled receptor 119; MGC119957;
Gene ID: 139760
mRNA Refseq: NM_178471
Protein Refseq: NP_848566
MIM: 300513
UniProt ID: Q8TDV5
Chromosome Location: Xq26.1
Pathway: 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; Synthesis, Secretion, and Inactivation of Glucagon-like Peptide-1 (GLP-1), organism-specific biosystem; Synthesis, Secretion, and Inactivation of Glucose-dependent Insulinotropic Polypeptide (GIP), organism-specific biosystem;
Function: G-protein coupled receptor activity; lipid 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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