All NIH-3T3 cell lines stably expressing EGFR and/or Mig6 were established by retroviral infections and pooled as described previously37 (link). NIH-3T3 and HEK-293T cells were purchased from ATCC and maintained in DMEM supplemented with either 10% bovine serum or 10% fetal bovine serum, respectively. PC9, HCC827 and A549 cells kindly provided by P. Janne (Dana-Farber Cancer Institute) were cultured in RPMI supplemented with 10% fetal bovine serum. Transient transfection was performed using X-tremeGene 9 (Roche) according to manufacture's instruction. Epidermal growth factor (EGF, Biosource) stimulations were performed using 25 ng/ml for 5 minutes unless noted in the text. All cell lines above except NIH-3T3 cells were authenticated by DNA finger printing analysis (STR analysis).
Protocol detail
Establishment and Characterization of EGFR and Mig6 Cell Lines
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.
293t cells
A549 cells
Bovine
Cancer
Cell lines
Egfr
Epidermal growth factor
Fetal bovine serum
Finger
Nih 3t3 cells
Retroviral infections
Serum
Transfection
Transient
Corresponding Organization : Samsung Medical Center
Other organizations : Boston University, National Cancer Center, Broad Institute
Top 5 similar protocols
Variable analysis
independent variables
- Stable expression of EGFR and/or Mig6 in NIH-3T3 cell lines (established by retroviral infections and pooled)
- Unspecified
- NIH-3T3 cells (without EGFR and/or Mig6 expression)
- HEK-293T cells
- PC9 cells
- HCC827 cells
- A549 cells
- DMEM supplemented with 10% bovine serum (for NIH-3T3 and HEK-293T cells)
- RPMI supplemented with 10% fetal bovine serum (for PC9, HCC827, and A549 cells)
- EGF stimulation at 25 ng/ml for 5 minutes
- Positive control: Unspecified
- Negative control: Unspecified
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!