安定細胞株製品 » Human Recombinant Gonadotropin-releasing Hormone Receptor Stable Cell Line
CHO-K1/GNRHR/Gα15 Stable Cell Line

Figure 1. Buserelin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/GNRHR/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with GNRHR agonist, buserelin. 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 buserelin (mean ± SEM, n = 3). The EC50 of buserelin on CHO-K1/GNRHR/Gα15 cells was 9.83 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 along a sigmoid curve.

CHO-K1/GNRHR/Gα15 Stable Cell Line

Figure 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with buserelin in CHO-K1/GNRHR/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/GNRHR/Gα15 cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of buserelin on CHO-K1/GNRHR/Gα15 cells was 81.97 nM.

CHO-K1/GNRHR/Gα15 Stable Cell Line

Recombinant CHO-K1 cells stably overexpress human gonadotropin releasing hormone receptor (GNRHR) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways.
M00426
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Product Introduction
Product Description Recombinant CHO-K1 cells stably overexpress human gonadotropin releasing hormone receptor (GNRHR) 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 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
Freeze Medium-DATA 45% culture medium, 45% FBS (Cat. No. 10099-141, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma)

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

    Figure 1. Buserelin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/GNRHR/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with GNRHR agonist, buserelin. 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 buserelin (mean ± SEM, n = 3). The EC50 of buserelin on CHO-K1/GNRHR/Gα15 cells was 9.83 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 along a sigmoid curve.

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

    Figure 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with buserelin in CHO-K1/GNRHR/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/GNRHR/Gα15 cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of buserelin on CHO-K1/GNRHR/Gα15 cells was 81.97 nM.


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