"GPBAR1" Related Products

Expand
Expand

Human GPBAR1/Clytin Stable Cell Line-HEK293

Cat.No.: CSC-RG0617
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 mitochondrially expressed photoproteins. Luminescent calcium assays offer several advantages over fluorescent calcium assays including:increased sensitivity and lack of interference from fluorescent compounds. These human GPBA receptor-expressing cells are made by stable transfection of HEK293 cells with clytin, receptor and clytin 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 GPBA receptor.
Background: The bile acid lithocolic acid acts as an agonist on the GPBA GPCR, also known as GPR131 or TGR5, to increase intracellular cAMP in cells that express the receptor. Bile acids and other GPBA ligands increase glucagon-like peptide-1 (GLP-1) secretion from intestinal cells. Although Type II diabetics have impaired ability to secrete GLP-1, their responsiveness to this compound, which enhances the effects of insulin, is normal. Thus GPBA agonists are of interest in developing therapeutics for diabetes. Interestingly, it has been suggested that the anti-diabetic effect of olive leaves could in part be mediated by agonist activity at GPBA receptors by oleanolic acid.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific silencing of GPBAR1.2. Protein expression: GPBAR1 in this cell line has been validated by immunocytochemical staining.
Application: Calcium Flux Assays; cAMP Assays
Sub-type: Bile Acid
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: GPBAR1 G protein-coupled bile acid receptor 1 [ Homo sapiens ]
Official Symbol: GPBAR1
Synonyms: GPBAR1; BG37; GPCR19; GPR131; M-BAR; TGR5; G protein-coupled bile acid receptor 1; G-protein coupled bile acid receptor 1; hBG37; hGPCR19; membrane bile acid receptor; G-protein coupled receptor GPCR19; membrane-type receptor for bile acids; G-protein coupled bile acid receptor BG37
Gene ID: 151306
mRNA Refseq: NM_001077191.1
Protein Refseq: NP_001070659.1
MIM: 610147
UniProt ID: Q8TDU6
Chromosome Location: 2q35
Pathway: Class A/1 (Rhodopsin-like receptors); GPCR ligand binding; Signal Transduction; Signal by GPCR; G alpha (s) signalling events; GPCR downstream signaling
Function: G-protein coupled receptor activity

Online Inquiry

Note: There will be extra charge for optional service!
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

Please input "biomart" as verification code. Please review Creative BioMart's privacy policy for more information


Price does not include shipping and packaging costs.Request quote for final price.

Price Inquiry

Welcome! For price inquiries, please feel free to contact us through the form below. We will get back to you as soon as possible.