Th1/Th2/Th17 CBA for cytokine production analysis

March 04, 2015

University of Nebraska Medical Center
Pharmacology and Experimental Neuroscience
Graduate Student

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Company:

BD Biosciences

Product Name:

Mouse Th1/Th2/Th17 Cytokine Kit

Catalog Number:

560485

The aim of my research is to determine the immune modulating capabilities of drug treatments in Parkinson's disease mouse models. I look at the ability of drug therapies to modulate T cell phenotypes following administration. I chose this product because it allows you to determine multiple cytokine concentrations in cell culture supernatant.

Experimental Design and Results Summary

Application

Cytokine Concentrations in Supernatants

Starting Material

CD4+ T cells

Protocol Overview

This CBA kit was used to detect the levels of IL-2, IL-4, IL-6, IFN-γ, TNF, IL-17A, and IL-10 in CD4+ cell culture supernatants following stimulation and drug treatment. CD4+ T cells were isolated from mice treated with drug, and were cultured. I isolated CD4+ T cells from both PBS and drug-treated mice. I cultured them, and stimulated with PMA and ionomycin to induce cytokine release, and then I collected the cell supernatants at varying time points to determine the amount of each cytokine within the Th1/Th2/Th17 kit. The supernatants were collected and detected using the fluorescent detection reagents in the CBA kit.  

Tips

Be very careful when using vacuum suction on the 96 well plates! The detection beads can be aspirated if the suction is too harsh!

Results Summary

I was able to successfully detect each of the cytokines within my cell supernatants. The standard curves were easily prepared and had fantastic R values, and the general protocol was very handy and easy to follow.

Additional Notes

None

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Summary

The Good

Easy to use and allows detection of multiple cytokines in a single sample.

The Bad

Expensive and need the software to be able to run it

The Bottom Line

I thought the kit was great, and gave me reliable results. The fact that you can run multiple samples and look for Th1/Th2/Th17 cytokines at the same time makes the expense worthwhile! The standards for normalization were easy to prepare as well! I highly recommend this product.

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