Product Description |
Recombinant CHO-K1 cells stably express glucagon-like peptide-1 receptor on the surface and the luciferase reporter gene incorporated into the biological signaling pathways will be activated upon the binding of GLP-1. The surface expression of GLP1R is validated by luminescence analysis. This stable cell line product is designed for cell-based functional assays of GLP-1 or analogues targeting GLP1R.
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Applications |
Functional assays for GLP1R
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Host Cell |
CHO-K1
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Size |
2 vials of frozen cells (>1X106per vial in 1 ml)
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Storage |
Store cells in liquid nitrogen immediately upon receipt. Thaw and recover cells within one year from the date received.
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Mycoplasma Status |
Negative. The mycoplasma test was performed with MycoAlert™ PLUS Mycoplasma Detection Kit (Cat. No. LT07-318, Lonza).
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Stability |
Stable through more than 15 passages with no significant changes in assay performance or expression profile.
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Culture Properties |
Adherent
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Freeze Medium |
95% complete growth medium, 5% DMSO (Cat. No. D2650, Sigma)
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Complete Growth Medium |
Ham's F-12K (Kaighn's) (Cat. No. 21127-022, Gibco), 10% FBS (Cat. No. 10099-141C, Gibco)
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Culture Medium |
Ham's F-12K (Kaighn's), 10% FBS, 500 μg/ml geneticin (Cat. No. 10131-035, Gibco), 250 μg/ml Hygromycin B (Cat. No. 10687010, Gibco)
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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.
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Figure 1: Functional evaluation of CHO-K1/CRE-CBLuc/GLP1R cells by GLP-1 (7-37).CHO-K1/CRE-CBLuc/GLP1R cells were plated and incubated at 37℃ for 16-18 hours prior to the addition of different concentrations of GLP-1 (7-37) (GenScript, RP12738-0.5). After 6 hours of incubation, detection reagent was added and the luminescence signal was measured by PHERAstar FSX Microplate Reader (BMG LABTECH). The relative light unit (RLU) were recorded and normalized to plot against the log of the cumulative doses of GLP-1 (7-37) (mean ± SD, n = 3). The EC50of GLP-1 (7-37) were 0.12 nM.
Notes:
EC50value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom) / (1+10^ ((LogEC50-X)*Hill Slope))
X is the logarithm of concentration. Y is the response.
Y is RLU and starts at Bottom and goes to Top along a sigmoid curve.
For research use only. Not intended for human and animal therapeutic or diagnostic use.
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