Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Super TopFlash (STF) assay

TF Tristan W. Fowler
TM Troy L. Mitchell
CJ Claudia Y. Janda
LX Liqin Xie
ST Shengjiang Tu
HC Hui Chen
HZ Haili Zhang
JY Jingjing Ye
BO Brian Ouyang
TY Tom Z. Yuan
SL Sung-Jin Lee
MN Maureen Newman
NT Nikita Tripuraneni
ER Erica S. Rego
DM Devin Mutha
AD Archana Dilip
MV Meghah Vuppalapaty
HB Helene Baribault
WY Wen-Chen Yeh
YL Yang Li
request Request a Protocol
ask Ask a question
Favorite

Wnt signaling activity was measured using HEK293 cells containing a luciferase gene controlled by a Wnt-responsive promoter (Super TopFlash reporter assay), as previously reported26. In brief, cells were seeded at a density of 10,000 per well in 96-well plates 24 h prior to treatment at the presence of 3 μM IWP2 to inhibit the production of endogenous Wnt ligands. Recombinant WNT3A (R&D systems) was used as a positive control. Cells were lysed with Luciferase Cell Culture Lysis Reagent (Promega), and activity was measured with the Luciferase Assay System (Promega), using vendor suggested procedures. Data were plotted as average −/+ standard deviation of triplicates and fitted by non-linear regression using Prism (GraphPad Software, San Diego, CA).

Copyright and License information: The Author(s) ©2021
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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