Luciferase Assay System

Protocol tips

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	After 24 hours after co-transfection determine luciferase. Remove medium and rinse cells in 1X PBS. Add 1X lysis reagent. Mix 20µl of cell lysate with 100µl of Luciferase Assay Reagent and measure the light produced.

Protocol tips
After 24 hours after co-transfection determine luciferase. Remove medium and rinse cells in 1X PBS. Add 1X lysis reagent. Mix 20µl of cell lysate with 100µl of Luciferase Assay Reagent and measure the light produced.

Publication protocol

Dual‐luciferase reporter assay was used to determine whether PTEN is a target gene of miR‐424‐3p. The pmirGLO vector was purchased from Promega (Madison, WI, USA). Recombinant vectors containing wild‐type (pmirGLO‐Wt‐PTEN 3′UTR) and mutant‐type (pmirGLO‐Mut‐PTEN 3′UTR) PTEN mRNA 3′UTR were constructed by Shanghai Biological Technology Company (Shanghai, China). miR‐424‐3p mimics or miR‐scramble were cotransfected with pmirGLO‐Wt‐PTEN 3′UTR or pmirGLO‐Mut‐PTEN 3′UTR into A549 and H460 cells, creating four test groups. Luciferase activity was determined 24 hours after cotransfection, using a luciferase assay kit (Promega, Madison, WI, USA) on a luminescence microplate reader (Berthold, Bad Wildbad, Germany).

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Manufacturer protocol

Download the product protocol from Promega for Luciferase Assay System below.

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