安定細胞株製品 » Human Recombinant Glucagon-like Peptide 1 Receptor Stable Cell Line
CHO-K1/GLP1R/Gα15 Stable Cell Line

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.

CHO-K1/GLP1R/Gα15 Stable Cell Line

CHO-K1/GLP1R/Gα15 Stable Cell Line

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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Overview
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

Product Introduction
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 Conditions
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)

Background
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.

Examples
  • CHO-K1/GLP1R/Gα15 Stable Cell Line
    • CHO-K1/GLP1R/Gα15 Stable Cell Line

    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.

  • CHO-K1/GLP1R/Gα15 Stable Cell Line
    • CHO-K1/GLP1R/Gα15 Stable Cell Line