| Product Description |
Recombinant HEK293 cells stably overexpress human glucagon-like peptide-1 (GLP1R) on the surface and contain high levels of G protein Gαs to couple with the receptor in downstream signaling pathways. |
| Applications |
FACS, Luciferase assay, cAMP accumulation assay. |
| Expressed Gene |
NCBI reference sequence NM_002062; no expressed tags |
| Target Protein |
NP_002053 |
| Host Cell |
HEK293 |
| Size |
Two vials of frozen cells (>1×106 per vial in 1 mL) |
| Storage |
Store cells in liquid nitrogen immediately upon receipt. Thaw and recover cells within one year from the date received. |
| Mycoplasma Status |
Negative. The mycoplasma test was performed with MycoAlert™ PLUS Mycoplasma Detection Kit (Cat. No. LT07-318, Lonza). |
| Stability |
Stable through more than 16 passages with no significant changes in assay performance or expression profile. |
| Culture Properties |
Adherent |
| Freeze Medium |
95% FBS (Cat. No. 10099-141, Life Technologies), 5% DMSO (Cat. No. D2650, Sigma) |
| Complete Growth Medium |
DMEM (Cat. No. 10566, Life Technologies), 10% FBS |
| Culture Medium |
DMEM, 10% FBS, 400 μg/ml Geneticin (Cat. No. 10131-035, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies) |
| Gene Synonyms |
GLP-1, GLP-1-R, GLP-1R; |
| Background |
GLP1R binds specifically the glucagon-like peptide-1 (GLP1) and has much lower affinity for related peptides such as the gastric inhibitory polypeptide and glucagon. GLP1R is known to be expressed in pancreatic beta cells. Activated GLP1R stimulates the adenylyl cyclase pathway which results in increased insulin synthesis and release of insulin. Consequently, GLP1R has been suggested as a potential target for the treatment of diabetes. GLP1R is also expressed in the brain where it is involved in the control of appetite. Furthermore, mice which over express GLP1R display improved memory and learning. |
Figure 1. GLP-1 (7-37)-induced luciferase expression in HEK293/CRE-Luc/GLP1R cells. After stimulation by agonist GLP-1 (7-37), the cells are determined with Fire-Lumi™ Luciferase Assay System (Cat. No. L00877C-100, Genscript) and the relative luminescence units (RLU) were recorded by plate reader (Pherastar, BMG). The RLU were plotted against the log of the cumulative doses of GLP-1 (7-37) (Mean ± SEM, n = 3). The EC50 of GLP-1 (7-37) on cells was 0.29 nM.
Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with GLP-1 (7-37) in HEK293/CRE-Luc/GLP1R cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in HEK293/CRE-Luc/GLP1R cells competitively bind with Europium Cryptate-labeled anti-cAMP monoclonal antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by plate reader (Varioskan, ThermoFisher). The EC50 of GLP-1 (7-37) on cells was 0.29 nM.
Figure 3. FACS analysis of cell surface expression of human GLP1R on HEK293/CRE-Luc/GLP1R cells. The HEK293/CRE-Luc/GLP1R cells (blue) and the negative control HEK293 cells (red) were probed using GLP-1-Fc fusion protein (GenScript) and FITC-conjugated goat anti-human IgG (H+L) (Cat. No. A01318, Genscript).
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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