Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

TOP/FOP reporter assay

SW Sihan Wang
YH Yi Han
JZ Jing Zhang
SY Shu Yang
ZF Zeng Fan
FS Feiling Song
LH Lijuan He
WY Wen Yue
YL Yanhua Li
XP Xuetao Pei
ask Ask a question
Favorite

Every 6 × 105 IEC-6 cells were transfected with a total of 1 μg Topflash (Addgene) or negative control Fopflash (Addgene) and 0.1μg of a renilla cocktail using the Neon Transfection System (Life Technologies) according to manufacturer's instructions. The conditions used were 1350 V, 30 ms, and 2-time pulses. A total of 4 × 104 transfected IEC-6 cells were cultured in the medium with or without Me6 in a 24-well plate. After 48 h, IEC-6 cells were collected and treated with a Dual-Glo Luciferase Assay System (Promega, E2920) according to the manufacturer's instructions. The luciferase activity was detected by SpectraMax of Molecular Devices.

Copyright and License information:  ©2020 The author(s)
This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A