目录产品 » HEK293/SBE-Luc2P
TGF-β/SMAD Signaling Pathway SBE Reporter-HEK293 Cell Line

Figure 1: TGF-β1, TGF-β2 or TGF-β3 induced concentration-dependent stimulation of intracellular luciferase accumulation in HEK293/SBE-Luc cells.
HEK293/SBE-Luc cells were plated and incubated at 37 ℃ for 16-18 hours prior to the addition of different concentrations of TGF-β1 (Cat. No. Z03411, GenScript), TGF-β2 (Cat. No. Z03429, GenScript) and TGF-β3. (Cat. No. Z03430, GenScript). After 6 hours of incubation, Fire-Lumi™ Reagent (Cat. No. L00877C, GenScript) 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-β1, TGF-β2, TGF-β3 on HEK293/SBE-Luc cells were 0.095 ng/ml, 0.096 ng/ml, 0.135 ng/ml, respectively.

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.

TGF-β/SMAD Signaling Pathway SBE Reporter-HEK293 Cell Line

Figure 2: M7824 induced blockade of intracellular luciferase accumulation triggered by TGF-β1, TGF-β2 or TGF-β3 in HEK293/SBE-Luc cells.
HEK293/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 (Cat. No. L00877C, GenScript) 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 HEK293/SBE-Luc cells were 0.0048 μg/ml (TGF-β1 activation), 1.221 μg/ml (TGF-β2 activation), 0.0005 μg/ml (TGF-β3 activation).

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.

TGF-β/SMAD Signaling Pathway SBE Reporter-HEK293 Cell Line

HEK293/SBE-Luc2P is designed for monitoring the TGF-β/SMAD signaling pathway activity and used to screen activators or inhibitors of TGF-β/SMAD signaling pathway.
M00903
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产品概述
Product Description Recombinant HEK293 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.
Applications Monitor TGF signaling pathway activity and screen for activators or inhibitors of the TGF/SMAD signaling pathway

产品简介
Host Cell HEK293
Quantity 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.
Mycoplasma Status Negative. The mycoplasma test was performed with MycoAlert™ PLUS Mycoplasma Detection Kit (Cat. No. LT07-318, Lonza).

培养条件
Culture Properties Adherent
Freeze Medium 95% complete growth medium, 5% DMSO (Cat. No. D2650, Sigma)
Complete Growth Medium DMEM (Cat. No. 10569-010, Gibco), 10% FBS (Cat. No. 10099-141C, Gibco)
Culture Medium DMEM, 10% FBS, 250 μg/ml geneticin (Cat. No.10131-035, Gibco)

技术背景
Gene Synonyms SBE reporter, TGFβ reporter, TGF beta reporter, SMAD reporter
Background The transforming growth factor beta (TGF-β) signaling pathway is involved in a diverse range of cellular processes such as differentiation, cell cycle arrest, and immune regulation. TGF-β signaling has been linked to cardiac disease, cancer, Alzheimer’s and other human diseases. TGF-β proteins bind to receptors on the cell surface, initiating a signaling cascade that leads to phosphorylation and activation of SMAD2 and SMAD3, which then form a complex with SMAD4. The SMAD complex then translocates to the nucleus and binds to the SMAD binding element (SBE) in the nucleus, leading to transcription and expression of TGF-β/ SMAD responsive genes.

图例
  • TGF-β/SMAD Signaling Pathway SBE Reporter-HEK293 Cell Line
    • TGF-β/SMAD Signaling Pathway SBE Reporter-HEK293 Cell Line

    Figure 1: TGF-β1, TGF-β2 or TGF-β3 induced concentration-dependent stimulation of intracellular luciferase accumulation in HEK293/SBE-Luc cells.
    HEK293/SBE-Luc cells were plated and incubated at 37 ℃ for 16-18 hours prior to the addition of different concentrations of TGF-β1 (Cat. No. Z03411, GenScript), TGF-β2 (Cat. No. Z03429, GenScript) and TGF-β3. (Cat. No. Z03430, GenScript). After 6 hours of incubation, Fire-Lumi™ Reagent (Cat. No. L00877C, GenScript) 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-β1, TGF-β2, TGF-β3 on HEK293/SBE-Luc cells were 0.095 ng/ml, 0.096 ng/ml, 0.135 ng/ml, respectively.

    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.

  • TGF-β/SMAD Signaling Pathway SBE Reporter-HEK293 Cell Line
    • TGF-β/SMAD Signaling Pathway SBE Reporter-HEK293 Cell Line

    Figure 2: M7824 induced blockade of intracellular luciferase accumulation triggered by TGF-β1, TGF-β2 or TGF-β3 in HEK293/SBE-Luc cells.
    HEK293/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 (Cat. No. L00877C, GenScript) 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 HEK293/SBE-Luc cells were 0.0048 μg/ml (TGF-β1 activation), 1.221 μg/ml (TGF-β2 activation), 0.0005 μg/ml (TGF-β3 activation).

    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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