Primer is superier to other version of ECL

November 04, 2014

Overall

Quality of Results

Ease-of-Optimization

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Duke University
Immunology
Postdoc

Company:

GE Healthcare

Product Name:

ECL Prime

Catalog Number:

RPN2232

I was trying to get a Western blot done for tissue expression of the trans gene from transgenic mice we created. I was having trouble with the ECL that we had in our lab which was ECL plus. Since we didn't know the tissue expression level of our transgene, we were looking for an ECL substrate which would be highly sensitive for detection of even small protein levels in tissues. I found out about ECL Prime from GE which claims higher sensitivity and better signal strength than ECl plus. I used if for the very first time and got very good results after doing certain optimization procedures like diluting the Primary as well as secondary antibody two times to its recommended concentration. I will now continue using this product and recommend it to others who wants even tiny protein to come up on film.

Experimental Design and Results Summary

Application

Western blot

Starting Material

Transgene expressing tissue samples

Protocol Overview

I diluted the primary and secondary two times more than the recommended concentration by manufacturer. The rest of the process was similar to that with other ECLs.

Tips

Dilute the primary as you might get high background. Washing is also very important after secondary incubation.

Results Summary

Results were excellent with this reagent. I was successful in finding the level of expression of my gene of interest. Although I had some bubble issues with a few samples, I got a clear image with many bands on the film.

Additional Notes

NA

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Summary

The Good

Highy sensitivity and signal stability

The Bad

Chance of high background

The Bottom Line

Product is very good for even low amount of protein. I used 20 ug but I guess even 10 ug would be sufficient. People should adjust amount according to their sample type and level of expression in vitro or in vivo.

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