| Product Description |
Recombinant CHO-K1 cells stably overexpress human growth hormone secretagogue receptor (GHSR) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways. |
| Applications |
Calcium mobilization assay, IP-One accumulation assay. |
| Expressed Gene |
NCBI reference sequence NM_198407; no expressed tags |
| Target Protein |
NP_940799 |
| 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, Gibco), 45% FBS (Cat. No. 10099-141, Gibco), 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, 500 μg/ml G418 (Cat. No. 10131-035, Gibco) |
| Gene Synonyms |
GHDP, GHS-R1a-1a, GHSR, ghrelin receptor |
| Background |
Ghrelin, a 28-amino-acid gut peptide is secreted from the stomach. It stimulates the release of growth hormone (GH) from the pituitary through GHS-R and plays important roles in energy homeostasis. In addition, ghrelin acts directly on the central nervous system to decrease sympathetic nerve activity. Ghrelin receptors (GHSRs) are concentrated in the hypothalamus-pituitary unit and may take the role of ghrelin in memory. GHSR is distributed in peripheral tissues, including the heart, lung, liver, kidney, pancreas, stomach, small and large intestines, adipose, and immune cells, suggesting that ghrelin has multiple functions in these tissues. GenScript's cloned human GHSR-expressing cell line was generated in CHO-K1 cell. |
Figure 1: Ibutamoren -induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/GHSR cells. The cells were loaded with Calcium-4 prior to being stimulated with a GHSR receptor agonist, Ibutamoren. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of Ibutamoren (Mean ± SD, n = 2). The EC50 of Ibutamoren on this cell was 14.28 nM.
Note:
1. EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom)/(1+10^((LogEC50-X)*HillSlope))
X is the logarithm of concentration.
Y is the response and starts at Bottom and goes to Top with a sigmoid shape.
2. Signal to Background Ratio (S/B) = Top/Bottom
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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