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| M00453 | |
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| Product Description | Recombinant CHO-K1 cells stably overexpress human prostaglandin D2 receptor 1 (DP1) on the surface and contain high levels of G protein Gαs 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. |
| 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) |
| Complete Growth Medium | Ham’s F-12K (Kaighn’s) (Cat. No. 21127, Life Technologies), 10% FBS |
| Freeze Medium-DATA | 45% culture medium, 45% FBS (Cat. No. 10099-141, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma) |
Figure 1. PGD2-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/DP1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with DP1 agonist, PGD2. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and normalized to plot against the log of the cumulative doses of PGD2 (mean ± SEM, n = 3). The EC50 of PGD2 on CHO-K1/DP1/Gα15 cells was 18.48 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 % stimulation of RFU and starts at Bottom and goes to Top along a sigmoid curve.
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Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with PGD2 in CHO-K1/DP1/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/DP1/Gα15 cells competitively bind with Europium Cryptate-labeled anti-cAMP monoclonal antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of PGD2 on CHO-K1/DP1/Gα15 cells was 9.49 nM. »
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
