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| M00434 | |
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| Product Description | Recombinant CHO-K1 cells stably overexpress human calcium sensing receptor (CASR) 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, 200 μg/ml Zeocin (Cat. No. R250-01, 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, Gibco), 10% DMSO (Cat. No. D2650, Sigma) |
Figure 1. CaCl2-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/CASR cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to being stimulated with agonist CaCl2. 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 CaCl2 (Mean ± SEM, n = 3). The EC50 of CaCl2 on this cell was 0.33 mM.
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 with a sigmoid shape.
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Figure 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with CaCl2 in CHO-K1/CASR cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/CASR cells competitively bind with Europium Cryptate-labeled anti-IP-One monoclonal antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Varioskan Lux, Thermo Fisher). The EC50 of CaCl2 on CHO-K1/CASR cells was 44.17 mM. »
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
