安定細胞株製品 » Human Recombinant MC5 Melanocortin Receptor Stable Cell Line
CHO-K1/MC5/Gα15 Stable Cell Line

Figure 1. NDP-α-MSH-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/MC5 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to being stimulated with agonist NDP-α-MSH. 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 NDP-α-MSH (Mean ± SEM, n = 3). The EC50 of NDP-α-MSH on this cell was 0.46 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.

CHO-K1/MC5/Gα15 Stable Cell Line

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with NDP-α-MSH in CHO-K1/Gα15/MC5 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/Gα15/MC5 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 (Varioskan LUX, Thermo Fisher). The EC50 of NDP-α-MSH on CHO-K1/Gα15/MC5 cells was 9.84 nM.

CHO-K1/MC5/Gα15 Stable Cell Line

Recombinant CHO-K1 cells stably overexpress human melanocortin 5 receptor (MC5R) on the surface and contain high levels of G protein Gαs to couple with the receptor in downstream signaling pathways.
M00243
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Product Introduction
Product Description Recombinant CHO-K1 cells stably overexpress human melanocortin 5 receptor (MC5R) 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 Conditions
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) (Cat. No. 21127, Life Technologies), 10% FBS (Cat. No. 10099-141, Life Technologies)
Freeze Medium-DATA 95% complete growth medium, 5% DMSO (Cat. No. D2650, Sigma)

Examples
  • CHO-K1/MC5/Gα15 Stable Cell Line
    • CHO-K1/MC5/Gα15 Stable Cell Line

    Figure 1. NDP-α-MSH-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/MC5 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to being stimulated with agonist NDP-α-MSH. 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 NDP-α-MSH (Mean ± SEM, n = 3). The EC50 of NDP-α-MSH on this cell was 0.46 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.

  • CHO-K1/MC5/Gα15 Stable Cell Line
    • CHO-K1/MC5/Gα15 Stable Cell Line

    Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with NDP-α-MSH in CHO-K1/Gα15/MC5 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/Gα15/MC5 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 (Varioskan LUX, Thermo Fisher). The EC50 of NDP-α-MSH on CHO-K1/Gα15/MC5 cells was 9.84 nM.


For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.