Western blotting was performed as previously described [49 (link)]. Primary antibodies were: rat anti-ATE1 (Merck Millipore; clone 6F11), mouse anti-GAPDH (Abcam; ab8245), mouse anti-actin (08691001, MP Biomedicals), mouse anti-tubulin (T9026, Merck), rabbit anti-ERK (#4695), rabbit anti-phosphorylated ERK (#4370), rabbit anti-phosphorylated S6K (Thr421/Ser424, #9204), rabbit anti-V5 (#13202) and rabbit anti-AXIN1 (#2087) (all from Cell Signaling Technology). HRP-conjugated secondary antibodies were purchased from Jackson ImmunoResearch and proteins were visualized by chemiluminescence (Image-Quant LAS400, GE Healthcare).
Protocol detail
Comprehensive Western Blotting Protocol
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.
Corresponding Organization :
Other organizations : Technion – Israel Institute of Technology, Centre de Biologie du Développement, Université de Toulouse, Centre National de la Recherche Scientifique, Discovery Institute, Sanford Burnham Prebys Medical Discovery Institute, Laboratoire de Recherche en Sciences Végétales, Institut de Pharmacologie et de Biologie Structurale, University of Pennsylvania, National Cancer Institute, Jewish General Hospital, McGill University
Top 5 similar protocols
Variable analysis
independent variables
- Primary antibodies: rat anti-ATE1, mouse anti-GAPDH, mouse anti-actin, mouse anti-tubulin, rabbit anti-ERK, rabbit anti-phosphorylated ERK, rabbit anti-phosphorylated S6K, rabbit anti-V5, and rabbit anti-AXIN1
- Protein expression and phosphorylation levels
- HRP-conjugated secondary antibodies
- Chemiluminescence (Image-Quant LAS400, GE Healthcare)
- Positive controls: not specified
- Negative controls: not specified
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!