Buffer works great for transcription factors and cytokines

Benaroya Research Institute
Immunology
Postdoc

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Company:

eBioscience

Product Name:

Foxp3 staining kit

Catalog Number:

00-5523-00

This product was used while running experiments looking at pathogenic T cells in a lung infection model. We were trying to distinguish Th17 and Treg cells in lung tissue. Using this product, we have successfully stained foxp3, rorgt and other difficult transcription factors. This buffer works even better if you stain overnight with your intracellular stains. I've also found that you can cut the buffer and perm wash down by half volume and it still works great! I chose this product because it was far superior to the competitor's buffer set in a side by side comparison.

Experimental Design and Results Summary

Application

Flow cytometry

Starting Material

Lung tissue

Protocol Overview

Homogenized lung, stain 4x10^6 cells for surface markers, transcription factors and cytokines. Assess by flow cytometry.

Tips

Leave in perm buffer with intracellular antibodies overnight

Results Summary

Was able to identify IL-17+RORgt+ CD4+ cells from whole Lung homogenate following infection. Also staining of foxp3 on eF450 (pacific blue) was greatly improved over other buffer systems.

Additional Notes

None

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Summary

The Good

Staining works great for foxp3

The Bad

Longer incubation times for best staining

The Bottom Line

Buy it! You won't regret it!

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