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| M00327 | |
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| Product Description | Recombinant CHO-K1 cells stably overexpress human complement C5a receptor 1 (C5AR1) 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. |
| Culture Medium | Ham’s F-12K (Kaighn’s), 10% FBS, 400 μg/ml Geneticin (Cat. No. 10131-035, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies) |
| Complete Growth Medium | Ham’s F-12K (Kaighn’s), 10% FBS |
| Freeze Medium-DATA | 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) |
Figure 1. W5Cha-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/C5a/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to being stimulated with agonist W5Cha. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (∆RFU) were recorded and plotted against the log of the cumulative doses of W5Cha (Mean ± SEM, n = 3). The EC50 of W5Cha on this cell was 0.11 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.
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