Figure 1. Glucagon-induced concentration-dependent stimulation of intracellular calcium mobilization in HEK293/GCGR/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist glucagon. 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 glucagon (Mean ± SEM, n = 3). The EC₅₀ of glucagon on this cell was 37.92 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 with a sigmoid shape.

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with glucagon in HEK293/GCGR/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in HEK293/GCGR/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 glucagon on HEK293/GCGR/Gα15 cells was 5.01 nM.

Figure 3. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with glucagon in HEK293/GCGR/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in HEK293/GCGR/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 glucagon on HEK293/GCGR/Gα15 cells was 0.60 μM.

HEK293/GCGR/Gα15 Stable Cell Line

Recombinant HEK293 cells stably overexpress human glucagon receptor (GCGR) on the surface and contain high levels of G protein Gαs to couple with the receptor in downstream signaling pathways.

產品編號	M00422
規格	2 vials 大單詢價
數量
價格	詢價


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產品簡介

Product Description	Recombinant HEK293 cells stably overexpress human glucagon receptor (GCGR) 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.

培養條件

Culture Medium	DMEM, 10% FBS, 300 μg/ml geneticin (Cat. No. 10131-035, Life Technologies), 50 μg/ml Hygromycin B (Cat. No. 10687010, Invitrogen)
Complete Growth Medium	DMEM, 10% FBS
Freeze Medium-DATA	50% DMEM (Cat. No. 10566, Life Technologies), 45% FBS (Cat. No. 10099-141, Gibco), 5% DMSO (Cat. No. D2650, Sigma)

圖例

Figure 1. Glucagon-induced concentration-dependent stimulation of intracellular calcium mobilization in HEK293/GCGR/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist glucagon. 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 glucagon (Mean ± SEM, n = 3). The EC₅₀ of glucagon on this cell was 37.92 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 with a sigmoid shape.

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with glucagon in HEK293/GCGR/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in HEK293/GCGR/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 glucagon on HEK293/GCGR/Gα15 cells was 5.01 nM.

Figure 3. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with glucagon in HEK293/GCGR/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in HEK293/GCGR/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 glucagon on HEK293/GCGR/Gα15 cells was 0.60 μM.

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

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HEK293/GCGR/Gα15 Stable Cell Line

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