Good Proliferation Dye For Human PBMCs

University College London
Infection and Immunity
PhD Student

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Company:

Thermo Fisher Scientific

Product Name:

Cell Proliferation Dye eFluor® 450 500 ug

Catalog Number:

65-0842-85

The aim of this study was to observe the proliferation of labeled target cells when co-cultured with unlabeled, HIV-infected donor cells. I chose this product as eBioscience by Thermo Fisher Scientific offers high-quality reagents at good prices.

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

Human PBMCs

Protocol Overview

Human PBMCs were stimulated with CD3/CD28 antibodies and then labelled with the proliferation dye according to manufacturer's instructions. Cells were surface stained, fixed and analysed by flow cytometry 5 days later.Proliferation dye staining:5e6 PBMCs were stained in 500ul HBSS with 5uM proliferation dye for 10 min at 37deg. Cells were washed in 5ml cRPMI and then cultured as normal in medium supplemented with IL-2.

Tips

Use serum/protein free medium for staining (PBS works as well). For shorter experiments (up to 3 days) you can use 2.5uM dye concentration.

Results Summary

As expected, we can observe good dilution of the proliferation dye in CD8+ T cell population, but not with CD4+ T cells.

DOI or PMID #

NA

Additional Notes

NA

Image Gallery

Summary

The Good

Good staining with long retention of signal

The Bad

None

The Bottom Line

I would recommend this reagent for flow cytometry.

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