| Product Description |
Recombinant CHO-K1 cells stably overexpress human 5-hydroxytryptamine receptor 2A (5-HT2A) on the surface and contain high levels of G protein Gαq and Gαi to couple with the receptor in downstream signaling pathways. |
| Applications |
Calcium flux assay |
| Expressed Gene |
NCBI reference sequence NM_000621; no expressed tags |
| Target Protein |
NP_000612 |
| 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. 10099-141, 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 Geneticin (Cat. No. 10131-035, Life Technologies) |
| Gene Synonyms |
HTR2, HTR2A |
| Background |
5-Hydroxytryptamine (5-HT, also known as serotonin) is synthesized in enterochromaffin cells in the intestine and in serotonergic nerve terminals. In the periphery, 5-HT mediates gastrointestinal motility, platelet aggregation, and contraction of blood vessels. Many functions of the central nervous system are influenced by 5-HT, including sleep, motor activity, sensory perception, arousal, and appetite. A family of 12 GPCRs and one ion channel mediate the biological effects of 5-HT (Hoyer et al., 1994). 5-HT2A which couples to Gq/11 is expressed throughout the central nervous system in the neocortex and olfactory tubercle. 5-HT2A receptor agonists may have important clinical value in the treatment of various disorders, such as depression, anxiety, bipolar disorder, and schizophrenia. GenScript's cloned human 5-HT2A–expressing cell line is generated in the CHO-K1 host. |
Figure 1. 5-HT-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/5-HT2A cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist 5-HT. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of 5-HT (Mean ± SEM, n = 3). The EC50 of 5-HT on this cell was 6.15 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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