目录产品 » 稳定细胞系 » Human Recombinant G-Protein Coupled Receptor 103 Stable Cell Line

Figure 1. QRFP26-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/GPR103/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist QRFP26 (26RFa). The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were plotted against the log of the cumulative doses of 26RFa (Mean ± SEM, n = 3). The EC₅₀ of 26RFa on this cell was 0.16 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 26RFa in CHO-K1/GPR103/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/GPR103/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 (Pherastar, BMG). The EC₅₀ of 26RFa on CHO-K1/GPR103/Gα15 cells was 0.23 μM.

CHO-K1/GPR103/Gα15 Stable Cell Line

GPR103 is known as an orphan G protein-coupled receptor with reported expression in brain, heart, kidney, adrenal gland, retina, and testis. GPR103 mRNA has been reported to be highly expressed in the superficial layers of the entire spinal cord and a high density of 26RFa binding sites was observed in the superficial layers of the dorsal horn. QRFP binds and activates the human GPR103, as well as mouse GPR103A and GPR103B, with nanomolar affinities in transfected cells. Therefore, the current experiments investigated the effects of QRFP administration in rats and the effects of a high fat diet on prepro-QRFP mRNA and GPR103 receptor mRNA levels. 26RFa and QRFP are endogenous ligands of GPR103. An immunohistochemical study revealed that GPR103-like immunoreactivity (LI) was observed in the superficial layers of spinal dorsal horn, that QRFP-LI was observed in the dorsal root ganglion and that intrathecal 26RFa suppressed the expression of Fos-LI induced by paw formalin injection in the superficial layers of the spinal dorsal horn.

产品编号	M00321
规格	2 vials 大单询价
数量
价格	询价


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产品简介

Product Description	Recombinant CHO-K1 cells stably overexpress human pyroglutamylated RFamide peptide receptor (QRFPR) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways.
Culture Properties	Adherent.
Stability	Stable through more than 16 passages with no significant changes in assay performance or expression profile.
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.

培养条件

Culture Medium	Ham’s F-12K (Kaighn’s), 10% FBS, 400 μg/ml Geneticin (Life Technologies), 100 μg/ml Hygromycin B (Life Technologies)
Complete Growth Medium	Ham’s F-12K (Kaighn’s), 10% FBS
Freeze Medium-DATA	45% Ham’s F-12K (Kaighn’s) (Life Technologies), 45% FBS (Life Technologies), 10% DMSO (Sigma)