HEK293 cells were grown in DMEM supplemented with 10% FBS and transfected with Fugene-HD Transfection Reagent (Roche), reporter constructs (pAhR-Luc/pTK-Renilla or pGud-Luc/pTK Renilla) as well transcription factor expression constructs (pStat1, pIrf9) as indicated. In some experiments, transfected cells were exposed to 500 IU/ml of mIFN-β (Bio X cell) or AhR inhibitor CH223191 (Sigma). For the measurement of AhR ligands in human serum, 15.000 HEK293 cells were plated in 96 well plates 24 h before transfection with pGud-Luc and pTK-Renilla. After 24 h transfected cells were incubated with DMEM supplemented with 10% of patient serum. Luciferase activity was analyzed 24 h later using Dual Luciferase Reporter System (Promega) and normalized to Renilla luciferase activity.
Protocol detail
Measurement of AhR Ligands in Human Serum
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Cells
Ch223191
Fugene
Hek293 cells
Human
Ligands
Luciferase
Patient
Promega
Renilla
Renilla luciferase
Serum
Transcription factor
Transfection
Corresponding Organization :
Other organizations : Brigham and Women's Hospital, Harvard University, Montreal Neurological Institute and Hospital, McGill University, University of Pennsylvania, Université de Montréal, Universidad de Sevilla, Weizmann Institute of Science, Broad Institute, Massachusetts Institute of Technology, University of Freiburg
Top 5 similar protocols
Protocol cited in 3 other protocols
Variable analysis
independent variables
- Transfection with reporter constructs (pAhR-Luc/pTK-Renilla or pGud-Luc/pTK Renilla)
- Transfection with transcription factor expression constructs (pStat1, pIrf9)
- Exposure to 500 IU/ml of mIFN-β (Bio X cell)
- Exposure to AhR inhibitor CH223191 (Sigma)
- Luciferase activity measured using Dual Luciferase Reporter System (Promega) and normalized to Renilla luciferase activity
- HEK293 cells grown in DMEM supplemented with 10% FBS
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!