Product Description |
Recombinant CHO-K1 cells
stably overexpress human glucagon-like peptide 1 receptor (GLP1R) and Gα15
subunit. Activated GLP1R stimulates the adenylyl cyclase pathway naturally.
While the co-expression of GLP1R and Gα15
subunit makes activated GLP1R also triggers PLC signaling and then stimulates
intracellular calcium mobilization. This cell line is recommended for screening
and functional validating agonists of GLP1R. |
Applications |
Screening and functional validation of agonists of
GLP1R |
Expressed Gene |
Codon optimized from NM_002062; no expressed tags |
Target Protein |
NP_002053 |
Host Cell |
CHO-K1/Gα15 |
Quantity |
2 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 |
45% Culture medium, 45% FBS (Cat. No. 10099-141,
Gibco), 10% (V/V) DMSO (Cat. No. D2650, Sigma) |
Complete Growth Medium |
Ham's F-12K (Kaighn's) Medium (Cat. No. 21127-022,
Gibco), 10% FBS (Cat. No. 10099-141, Gibco) |
Culture Medium |
Ham's F-12K (Kaighn's) Medium (Cat. No. 21127-022,
Gibco), 10% FBS (Cat. No. 10099-141, Gibco), 200 μg/ml Zeocin (Cat. No. R250-01, Life
Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Invitrogen) |
Gene Synonyms |
GLP-1R |
Background |
GLP1R binds specifically
the glucagon-like peptide-1 (GLP-1) and has much a 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 overexpress
GLP1R display improved memory and learning. |
Figure 1: GLP-1 (7-37)-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/GLP1R/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular DEVICES) prior to stimulation with GLP1R agonist, Glucagon-Like Peptide (GLP) I (7-37) (Cat. No. RP12738, GenScript). The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescence units (RFU) recorded and normalized to plot against the log of the cumulative doses of GLP-1 (7-37) (Mean ± SD, n = 2). The EC50 of GLP-1 (7-37) on CHO-K1/GLP1R/Gα15 cells was 0.50 µM.
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 RFU and starts at Bottom and goes to Top along a sigmoid curve.
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