| 询价 | |
| M00902 | |
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| Product Description |
Recombinant HT1080 SBE reporter
stable cell line expresses luciferase in response to activation of TGFβ/SMAD signaling
pathway. This stable cell line product is designed for monitor TGF signaling
pathway activity and screen for activators or inhibitors of the TGF/SMAD
signaling pathway. |
| Culture Properties |
Adherent |
| Storage |
Store
cells in liquid nitrogen immediately upon receipt. Thaw and recover cells
within one year from the date received. |
| Culture Medium |
MEM,
10% FBS, 500 μg/ml geneticin (Cat. No. 10131-035, Gibco) |
| Complete Growth Medium |
MEM
(Cat. No. 32561-037, Gibco), 10% FBS (Cat. No. 10099-141C, Gibco) |
| Freeze Medium-DATA |
95%
complete growth medium, 5% DMSO (Cat. No. D2650, Sigma) |
Figure 1: Functional evaluation of HT1080/SBE-Luc cells by TGF-β1, TGF-β2 and TGF-β3.
HT1080/SBE-Luc cells were plated and incubated at 37 ℃ for 16-18 hours prior to the addition of different concentrations of TGF-β1 (GenScript, Z03411), TGF-β2 (GenScript, Z03429) and TGF-β3. (GenScript, Z03430). After 6 hours of incubation, Fire-Lumi™ Reagent (GenScript, L00877C) was added and the luminescence signal was measured by Thermo Scientific Varioskan LUX. The relative light unit (RLU) were recorded and normalized to plot against the log of the cumulative doses of TGF-β (mean ± SD, n = 2). The EC50 of TGF-β on HT1080/SBE-Luc cells were 0.096 ng/ml (TGF-β1), 0.058 ng/ml (TGF-β2), 0.075 ng/ml (TGF-β3). »
Figure 2: Functional evaluation of HT1080/SBE-Luc cells by M7824.
HT1080/SBE-Luc cells were plated and incubated at 37 ℃ for 16-18 hours. Mix TGF-β1 (0.3 ng/ml), TGF-β2 (0.3 ng/ml) or TGF-β3 (0.3 ng/ml) with different concentrations of M7824 for 30 min. After 30 min of incubation, the mixture was added into the 384-well plate and incubated at 37℃ for 6 hours. After incubation, Fire-Lumi™ Reagent (GenScript, L00877C) was added and luminescence measured by Thermo Scientific Varioskan LUX. The relative light unit (RLU) were recorded and normalized to plot against the log of the cumulative doses of TGF-β (mean ± SD, n = 2). The IC50 of M7824 on HT1080/SBE-Luc cells were 0.02 μg/ml (TGF-β1), 2.58 μg/ml (TGF-β2), 0.002 μg/ml (TGF-β3).
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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