ELISA Made Easy

June 30, 2016

University of Idaho
Biological Sciences
Graduate student

Overall

Quality of Results

Ease-of-Optimization

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Company:

eBioscience

Product Name:

Mouse TNF alpha ELISA Ready-SET-Go!

Catalog Number:

88-7324

I am looking at characterizing respiratory viral infections in mice.  My goal for this experiment is to determine the cytokine and chemokine profiles for these infections. I used this specific product because my lab had it on hand and was readily available.

Experimental Design and Results Summary

Application

Determining cytokine concentrations

Starting Material

Mouse bronchoalveolar lavage fluid

Protocol Overview

Apply detection antibody to the plate and incubate overnight. Wash five times and then block wells. Then wash five more times and add your standards to the appropriate wells and your samples to the other wells and incubate overnight. Wash five more times and add detection antibody and incubate for one hour. Wash another five times and add avidin/HRP. Incubate for 30 minutes. Wash seven times now and then add your substrate to the wells. After 15 minutes, add the stop solution (I used H3PO4). Read plate.

Tips

Soaking the washes for one minute does help the wash steps. Incubating overnight increases your readings.

Results Summary

After running the TNF alpha standard I generate a concentration curve. I can then apply my samples to the curve and determine the TNF alpha concentration in my samples.

Additional Notes

None

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Summary

The Good

It's pretty easy to use and the protocol is straightforward.

The Bad

I haven't run into any issues with this product so far.

The Bottom Line

This is a good way of determining your TNF alpha concentrations. I do two overnight incubations so it takes some time to run, but this is optional. i would recommend this product for others.

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