Moderate Protein Transport Inhibitor

October 24, 2018

BlueskyReddit
Johns Hopkins School of Medicine
Pharmacology and Oncology
Graduate Student

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Company:

BD Biosciences

Product Name:

Protein Transport Inhibitor (Containing Brefeldin A)

Catalog Number:

555029

As an immunology lab, we routinely assess intracellular cytokine production from T cells and other immune cells. We stimulate cells using anti-CD3/28 to induce cytokine production at moderate levels; therefore, we use this moderate protein transport inhibitor to block secretion in order to analyze cytokines using flow cytometry.

Experimental Design and Results Summary

Application

Analyze intracellular cytokine production

Starting Material

T cells or splenocytes

Protocol Overview

Obtain single-cell suspension of cells. Stimulate cells using appropriate stimulation with 1000x GolgiPlug. Stimulate 10–14 hours with this inhibitor since it does not lead to massive death.

Tips

Remove the inhibitor from the fridge long before use since it's frozen in DMSO.

Results Summary

We are able to detect robust IFNg secretion from CD8 T cells in conjunction with proliferation of cells.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Easy to use always at 1000x for most stimulation conditions. Gentle on T cells without causing too much cell death.

The Bad

Takes a while to thaw after removal from fridge.

The Bottom Line

Convenient way to block transport of cytokines. Ready-made mixture is always consistent, which is important for consistent results.

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