| Product Description |
Recombinant CHO-K1 cells stably overexpress human neuropeptide S receptor 1 (NPSR1) isoform B on the surface and contain high levels of G protein Gαs and Gαq to couple with the receptor in downstream signaling pathways. |
| Applications |
Calcium mobilization assay and IP-One assay. |
| Expressed Gene |
NCBI reference sequence NM_207173; no expressed tags |
| Target Protein |
NP_997056 |
| 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, 200 μg/ml Zeocin (Cat. No. R250-01, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies) |
| Gene Synonyms |
NPSR1, ASRT2, GPR154, GPRA, NPSR, PGR14, VRR1 |
| Background |
The NPS receptor is a typical GPCR, also known as GPR154, vasopressin-receptor related receptor 1 (VRR1), or GPRA. NPSR was found mainly expressed in the central nervous system of rats by using in-situ hybridization. NPS receptor mRNA is widely distributed in many brain areas with high expression levels in cortex, hypothalamus, amygdala and multiple midline thalamic nuclei. Many of these areas have been functionally associated with arousal and processing of emotional behavior. In 2004, the NPS receptor was identified as an asthma susceptibility gene in a genome wide screen in Finnish and Canadian patients. The study showed that a number of polymorphic variants of the NPS receptor exist in human and that particular sets of these variants (haplotypes) are associated with an increased risk of asthma and possibly allergic diseases characterized by high IgE serum levels. A carboxy-terminal splice variant of human NPS receptor was found to be over-expressed in asthmatic airway tissue. |
Figure 1. Neuropeptide S-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/NPS1b/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist neuropeptide S. 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 neuropeptide S (Mean ± SEM, n = 3). The EC50 of neuropeptide S on CHO-K1/NPS1b/Gα15 cell was 0.20 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 IP-One accumulation upon treatment with neuropeptide S on CHO-K1/NPS1b/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/NPS1b/Gα15 cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Varioskan, Thermo). The EC50 of neuropeptide S on CHO-K1/NPS1b/Gα15 cells was 0.10 μM.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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