Good ECL Western Blotting Reagent From ThermoFisher

Stanford University
Biochemistry
Senior Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

ThermoFisher Scientific

Product Name:

Pierce ECL Western Blotting Substrate

Catalog Number:

32106

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).

Experimental Design and Results Summary

Application

Western blotting

Starting Material

RKO whole cell lysate

Tips

5-10 sec incubation is more than enough.

Results Summary

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.

DOI or PMID #

N/A

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Summary

The Good

Super easy to use.

The Bad

None.

The Bottom Line

Good for moderate to high expressed genes (at protein level).

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