Figure 1. Orexin A-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OX2 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist orexin A. 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 orexin A (Mean ± SD, n = 2). The EC50 of orexin A on this cell was 1.3 nM.
Notes:
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
Y is RFU and starts at Bottom and goes to Top with a sigmoid shape.
CHO-K1/OX2 Stable Cell Line
Hypocretin (orexin) receptor 2, also known as OX2, is a human protein encoded by the HCRTR2 gene. Orexin A and orexin B are neuropeptides originally identified as endogenous ligands for OX2. Orexin neuropeptides are produced by a small group of neurons in the lateral hypothalamic and perifornical areas, a region that is classically implicated with the control of mammalian feeding behavior. Orexin neurons that project throughout the central nervous system to nuclei are known to be important in the control of feeding, sleep-wakefulness, neuroendocrine homeostasis, and autonomic regulation.
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