| 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.
|
| Applications |
Functional assays for
GLP1R
|
| Host Cell |
CHO-K1
|
| Size |
2 vials of frozen
cells (>1X106 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 15 passages with no significant changes in assay performance or expression
profile.
|
| Culture Properties |
Adherent
|
| Freeze Medium |
95%
complete growth medium, 5% DMSO (Cat. No. D2650, Sigma)
|
| Complete Growth Medium |
Ham's
F-12K (Kaighn's) (Cat. No. 21127-022, Gibco), 10% FBS (Cat. No. 10099-141C,
Gibco)
|
| 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)
|
| 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: 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 EC50 of GLP-1 (7-37) were 0.12 nM.
Notes:
EC50 value 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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