ThermoFisher Scientific
SuperSignal West Pico PLUS Chemiluminescent Substrate
34580
Info: Supplier Page
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 a WNT pathway target gene).
Western blotting
RKO whole cell lysate
Good for low expressed genes (at protein level).
25 µg of whole cell lysates from RKO (human colorectal cancer) cell lines were resolved on a 4-12% Bis-Tris gel, proteins transferred onto a nitrocellulose membrane, and immunoblotted for AXIN2 antibody. The membrane was incubated for 5 sec with the Pico PLUS reagent and the signal was captured on a HyBlot CL Autoradiography Film. AXIN2 is elevated in APC mutant background and better detected with the Pico PLUS reagent compared to the regular ECL substrate.
N/A
A good option if regular ECL fails in detection step.
None.
Good option for low expressed genes (at protein level).