Luciferase Assay System

Reporter gene assay β-galactosidase substrates - C2C12

Experiment
Reporter gene assay β-galactosidase substrates - C2C12
Product
Luciferase Assay System from Promega
Manufacturer
Promega

Protocol tips

Protocol tips
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

The FLAG-SIRT1 and FLAG-SIRT1 H355A genes were kindly provided by Professor Soo-Jong Um at Sejong University, and the myogenin- and creatine kinase promoter-luciferase reporter genes were provided by Professor Jeong-Ho Hong at Korea University. The mitsugumin 53 (MG53) promoter-luciferase reporter gene was previously generated (40). Gene transfection was performed using electroporation according to the protocol of electroporator MP-100 (Invitrogen) for C2C12 myoblasts. Luciferase activity was measured by using the luciferase assay system (Promega) using a Luminoskan Ascent (Thermo Labsystems). We normalized the luciferase activity to the activity of coexpressed β-galactosidase. To calculate the relative luciferase activity, we divided each normalized luciferase activity by that of the empty vector-transfected cells.

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Papers

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Paper title
Mitochondrial Complex I Deficiency Enhances Skeletal Myogenesis but Impairs Insulin Signaling through SIRT1 Inactivation
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Manufacturer protocol

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

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