| Product Description |
Human
Recombinant HEK293/SRE-Luc Stable Cell Line stably overexpress the luciferase
reporter (Luc) gene. This stable cell line product is designed for measuring the
function of EGF or anti-EGFR.
|
| Applications |
The HEK293/SRE-Luc Stable cell line is designed for
measuring the potency of EGF or anti-EGFR, and monitoring MAPK/ERK signaling
pathway activity.
|
| Expressed Gene |
Luciferase Reporter
|
| Host Cell |
HEK293
|
| Quantity |
2 vials of frozen cells (>1X106 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. # LT07-318, Lonza).
|
| Stability |
Stable through more than 15 passages with no
significant changes in assay performance or expression profile.
|
| Culture Properties |
Adherent
|
| Freeze Medium |
90%FBS (Cat. No. 10091-148, Gibco), 10% DMSO (Cat. No.
D2650, Sigma)
|
| Complete Growth Medium |
DMEM (Cat. # 10569-010, Gibco), 10% FBS (Cat. # 10091-148, Gibco)
|
| Culture Medium |
DMEM, 10% FBS, 300 μg/ml Geneticin (Cat. # 10131-035,
Gibco)
|
| Background |
Luciferase reporter assays are uniquely suited
for high throughput screening due to their inherently high sensitivity, wide
dynamic range and low susceptibility to compound interference. Here, we have
incorporated the technical advancement of luciferase reporter genes and vectors
to build an improved response element luciferase construct for MAPK/ERK
signaling pathway.
The MAPK/ERK signaling pathway
is a major participant in the regulation of cell growth and differentiation. It
can be activated by various extracellular stimuli including mitogens, growth
factors, and cytokines. Upon stimulation, MEK1/2 phosphorylate and activate
ERK1/2. The activated ERK translocates to the nucleus where it phosphorylates and
activates transcription factors. The TCFs (Ternary Complex Factors), including
Elk1, are among the best-characterized transcription factor
substrates of ERK. When phosphorylated by ERK, Elk1 forms a complex with the
response factor and binds to its response element, resulting in the expression
of numerous mitogen-inducible genes. |
Figure 1: Functional evaluation of HEK293/SRE-Luc cells by EGF. HEK293/SRE-Luc cells were plated and incubated at 37 ℃ for 16-18 hours prior to the addition of different concentrations of EGF (GenScript, Z02691). After 6 hours of incubation, Fire-LumiTM Reagent (GenScript, L00877) was added and luminescence measured by Thermo Scientific Varioskan LUX.
Figure 2: Functional evaluation of HEK293/SRE-Luc cells by Cetuximab. HEK293/SRE-Luc cells were plated and incubated at 37 ℃ for 16-18 hours prior to the addition of EGF (0.03 µg/ml, GenScript). After 30 min of incubation, different concentrations of Cetuximab were added into the 384-well plate and incubated at 37℃ for 6 hours. After 6 hours of incubation, Fire-LumiTM Reagent (GenScript, M00877) was added and luminescence measured by Thermo Scientific Varioskan LUX.
Figure 3: Functional evaluation of HEK293/SRE-Luc cells by FGF. HEK293/SRE-Luc cells were plated and incubated at 37 ℃ for 16-18 hours prior to the addition of different concentrations of FGF (GenScript, Z03116-50). After 6 hours of incubation, Fire-LumiTM Reagent (GenScript, L00877) was added and luminescence measured by Thermo Scientific Varioskan LUX.
For
research use only. Not intended for human and animal therapeutic or
diagnostic use.
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