ThermoFisher Scientific
SuperSignal West Femto Maximum Sensitivity Substrate
34095
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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 pathway component).
Western blotting
HAP1 whole cell lysate
Use this if Pico PLUS doesn't work.
25 µg of whole cell lysates from HAP1 (a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia) cell lines were resolved on a 4-12% Bis-Tris gel, proteins transferred onto a nitrocellulose membrane, and immunoblotted for APC antibody. The membrane was incubated for 5 sec with the Femto Western Blotting substrate and the signal was captured on a HyBlot CL Autoradiography Film. APC gene is expressed low at protein level in this cell line.
N/A
Works for the poorly expressed gene (at protein level).
The signal to noise ratio can be poor if exposure is long.
I use this as my last option if ECL Pico PLUS fails or is poor in detecting a signal.