Excellent Antibody For Staining of FoxP3+ Regulatory T Cells

September 24, 2015

Neurology
BWH
Postdoc

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Company:

eBioscience

Product Name:

Anti-Mouse/Rat Foxp3 FITC

Catalog Number:

11-5773-82

Info: View Product View Product Specs

Browse Similar Products: Foxp3 Antibodies

Our laboratory is interested in induction of tolerance in autoimmune diseases. Tolerance induction is often associated with induction of FoxP3+ regulatory T cells (Treg). Foxp3 is a member of the forkhead/winged-helix family of transcriptional regulators and is highly expressed by both, natural and in vitro induced Treg. This antibody is routinely used by our laboratory to identify FoxP3+ Treg.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse spleen cells

Primary Incubation

30min, 4'C, 1/200 dilution in FoxP3+ staining buffer (also eBiosciences) 50ul per sample

Blocking Agent

Fc block (anti CD16/CD32)

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

FACS LSRII

Results Summary

Spleen cells were gated on live CD4+ T cells and analyzed for expression of CD25 and FoxP3. FoxP3+CD25 high cells are highly suppressive regulatory T cells. FoxP3+CD25-/low cells are also regulatory T cells but with less suppressive capacity.

Additional Notes

Staining was performed according to the protocol provided by eBiosciences. First extracellular staining of surface markers (here CD4 and CD25) then fixation, permeabilization and intracellular staining using Foxp3 / Transcription Factor Staining Buffer Set (eBiosciences # 00-5523). These buffers also work great for detection of intracellular cytokines.

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Summary

The Good

Bright and specific staining

The Bad

Oobviously this product is unsuited for sorting of live cells as fixation and permeabilization are required. The use of FoxP3-gfp reporter mice is necessary for sorting live FoxP3+ T cells

The Bottom Line

Great product!

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