| Product Description |
Recombinant CHO-K1 cells stably overexpress human neuropeptide Y receptor Y4 (NPY4) on the surface and contain high levels of G protein Gαi and Gαq to couple with the receptor in downstream signaling pathways. |
| Applications |
Calcium mobilization assay and cAMP accumulation assay. |
| Expressed Gene |
NCBI reference sequence NM_005972; no expressed tags |
| Target Protein |
NP_005963 |
| 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. 10099-14, 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 G418 (Cat. No. 10131-035, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies) |
| Gene Synonyms |
NPY4R, NPY4-R, PP1, PPYR1, Y4 |
| Background |
The NPY family consists of three 36-amino acid peptides, neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP), which bind to the NPY receptors. At present five distinct NPY receptors, Y1, Y2, Y4, Y5, and Y6, have been established by receptor cloning studies and all of them are Gi-coupled GPCRs. Activation of NPY receptors mediate a variety physiological effects including stimulation of food intake, inhibition of anxiety in the CNS, presynaptic inhibition of neurotransmitter release in the CNS and periphery, modulation of circadian rhythm, release of pituitary hormones, modulation of hippocampal activity, pain transmission, vasoconstriction, inhibition of insulin release and modulation of renal function. Y4 receptor preferentially binds PP, with significant binding to NPY and PYY. |
Figure 1. Pancreatic polypeptide-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/NPY4/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist pancreatic polypeptide. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of pancreatic polypeptide (Mean ± SEM, n = 3). The EC50 of pancreatic polypeptide on CHO-K1/NPY4/Gα15 cell was 95.03 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 RFU and starts at Bottom and goes to Top with a sigmoid shape.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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