| Product Description |
Recombinant CHO-K1 cells stably overexpress human glucagon receptor (GCGR) on the surface and contain high levels of G protein Gαs to couple with the receptor in downstream signaling pathways. |
| Applications |
FACS, calcium mobilization assay, IP-One assay and cAMP accumulation assay. |
| Expressed Gene |
NCBI reference sequence NM_000160; no expressed tags |
| Target Protein |
NP_000151 |
| Host Cell |
CHO-K1 |
| 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 |
45% Ham’s F-12K (Kaighn’s) (Cat. No. 21127, Life Technologies), 45% FBS (Cat. No. 10099-141, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma) |
| Complete Growth Medium |
Ham’s F-12K (Kaighn’s), 10% FBS |
| Culture Medium |
Ham’s F-12K (Kaighn’s), 10% FBS, 400 μg/ml geneticin (Cat. No. 10131-035, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies) |
| Gene Synonyms |
GGR, GL-R |
| Background |
Glucagon regulates blood glucose via control of hepatic glycolgenolysis and gluconeogenesis and via regulation of insulin release from the β cell. Pharmacological administration of glucagon increases blood glucose in normal and diabetic subjects, and produces positive inotropic and chronotropic cardiovascular effects, relaxation of smooth muscle in the gastrointestinal tract and stimulation of growth hormone secretion. The actions of glucagon are mediated via a single adenylate cyclase-coupled glucagon receptor that also couples to the phospholipase C inositol phosphate (PLC-IP) pathway leading to Ca2+ release from intracellular stores. |
Figure 1. Glucagon-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/GCGR/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist glucagon. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of glucagon (Mean ± SEM, n = 3). The EC50 of glucagon on this cell was 0.68 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 with a sigmoid shape.
Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with glucagon in CHO-K1/GCGR/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/GCGR/Gα15 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 (Pherastar, BMG). The EC50 of glucagon on CHO-K1/GCGR/Gα15 cells was 7.24 nM.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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