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CHO-K1/Gα15/AMY2 Stable Cell Line

Recombinant CHO-K1 cells stably overexpress human receptor activity modifying protein 2 (RAMP2) and calcitonin receptor (CTR) on the surface. Amylin receptors (AMYR) are a class of special protein heterodimers, composed of a CTR combined with one of three homologous receptor activity - modifying proteins (RAMP1, RAMP2, RAMP3), forming three subtypes (AMY1, AMY2, AMY3). This cell contain high levels of G protein Gαs to couple with the receptor in downstream signaling pathways.
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M00558

产品简介
Product Description Recombinant CHO-K1 cells stably overexpress human receptor activity modifying protein 2 (RAMP2) and calcitonin receptor (CTR) on the surface. Amylin receptors (AMYR) are a class of special protein heterodimers, composed of a CTR combined with one of three homologous receptor activity - modifying proteins (RAMP1, RAMP2, RAMP3), forming three subtypes (AMY1, AMY2, AMY3). This cell 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.
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培养条件
Culture Medium Ham’s F-12K (Kaighn’s), 10% FBS, 200 μg/ml Zeocin (Cat. No. R250-01, 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)
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图例
CHO-K1/Gα15/AMY2 Stable Cell Line

Figure 1. Amylin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/AMY2 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with AMY2 agonist, amylin. 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 amylin (mean ± SEM, n = 3). The EC50 of amylin on CHO-K1/Gα15/AMY2 cells was 51.69 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. »

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