ThermoFisher Scientific
Pierce ECL Western Blotting Substrate
32106
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The WNT pathway is essential for embryonic development and adult tissue homeostasis. De-regulation of this pathway results in cancers of the stomach, intestine, breast, etc. I am interested in studying the molecular mechanism of oncogenic WNT signal transduction to identify potential targets for therapeutic intervention of colon cancer. This project involves monitoring mRNA and protein levels of pathway components in the presence of WNT ligand (physiological) or in the background of APC gene truncation (oncogenic). This experiment shows the signal sensitivity of an ECL reagent after immunoblotting for alpha-Tubulin (a loading control).
Western blotting
RKO whole cell lysate
5-10 sec incubation is more than enough.
25 µg of whole cell lysates from RKO (human colorectal cancer) cell line were resolved on a 4-12% Bis-Tris gel, proteins transferred onto a nitrocellulose membrane, and immunoblotted for alpha-Tubulin antibody. The membrane was incubated for 5 sec with the Pierce ECL Western blotting substrate and the signal was captured on a HyBlot CL Autoradiography Film.
N/A
Super easy to use.
None.
Good for moderate to high expressed genes (at protein level).