The 3′ UTR sequence of MEF2C containing wild-type or mutant binding sites was cloned into the pmirGLO luciferase vector. Then, the MEF2C 3′ UTR WT or MUT and miR-524-5p mimics or negative control were cotransfected using Lipofectamine 3000. After 72 h, the cell lysate was centrifuged at 12,000 rpm for 5 min to perform the Dual-Lumi Luciferase Assay (Beyotime Biotechnology, Shanghai, China). Luciferase activity was measured by an EnVision Multifunctional Microplate Reader (PerkinElmer, Germany).
Dual lumi luciferase assay
Manufactured by Beyotime
Sourced in China
The Dual-Lumi Luciferase Assay is a lab equipment product that measures the activity of two luciferase reporter enzymes simultaneously. It utilizes a dual-emission detection system to quantify the luminescent signals generated by the two luciferase reporters. The core function of this product is to enable the concurrent analysis of two distinct biological processes or gene expression events within the same sample.
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Lab products found in correlation
2 protocols using dual lumi luciferase assay
1
Luciferase Assay for MEF2C 3'UTR Regulation
2
Luciferase Assay for MEF2C 3'UTR
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The MEF2C 3' UTR sequence containing wild-type and mutant binding sites was cloned into the pmirGLO luciferase vector. After the MEF2C 3'UTR WT (or MUT) were co-transfected with miR-524-5p mimic or negative control by Lipofectamine 3000 respectively. After 72 h, the cells were lysed and centrifuged at 12,000 rpm for 5 min to perform Dual-Lumi Luciferase Assay (Beyotime Biotechnology, Shanghai, China). The luciferase activity of the cells was determined by EnVision Multifunctional Microplate Reader (PerkinElmer, Germany). pmirGLO vector transfected into cells served as internal control.
Cell Counting Kit-8 (CCK-8) assay 100 μL of cell suspension containing 1×104 cells was added in each well of the 96-well plate. At 6 hrs, 24 hrs, 48 hrs, 72 hrs, and 96 hrs, 10 μL of CCK-8 reagent (Beyotime Biotechnology, Shanghai, China) was supplied, respectively. After cell culture for 2 hrs, the absorbance value of each well at 450 nm wavelength was measured by a microplate reader for plotting a growth curve.
Cell Counting Kit-8 (CCK-8) assay 100 μL of cell suspension containing 1×104 cells was added in each well of the 96-well plate. At 6 hrs, 24 hrs, 48 hrs, 72 hrs, and 96 hrs, 10 μL of CCK-8 reagent (Beyotime Biotechnology, Shanghai, China) was supplied, respectively. After cell culture for 2 hrs, the absorbance value of each well at 450 nm wavelength was measured by a microplate reader for plotting a growth curve.
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