Monoclonal FOXP3-APC Anti-Mouse Antibody for Flow Cytometry

Vaccine Immunology Laboratory
National Institute of Immunology, India
Postdoctoral Researcher

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Company:

Invitrogen

Product Name:

FOXP3-APC monoclonal antibody for flow cytometry

Catalog Number:

17-5773-80

Regulatory T cells are a specialized subset of CD4+ T cells involved in suppression of immune responses and identified by the expression of CD4+CD25+FOXP3+ markers. To detect the percent of regulatory T cells present in splenocytes after vaccine exposure, we have used this antibody in our antibody cocktail for multi-color flow cytometry.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mice splenocytes

Primary Incubation

1:20 dilution in perm buffer, 4°C, 1 hour (1–2 million cells)

Blocking Agent

Fc-block and fixation of cells for 30 minutes using intracellular fixation & permeabilization bufffer from eBioscience

Secondary Incubation

None

Tertiary Incubation

None

Detection

Flow cytometry

Results Summary

Live CD4+ T cells were gated for the expression of CD25 and FOXP3 on dot plot. Double positive population (CD25+FOXP3+) is detected as regulatory T cells. Clear cloud for FOXP3+ cells were observed.

DOI or PMID #

N/A

Additional Notes

Always use the antibody with intracellular fixation & permeabilization buffer.

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Summary

The Good

Good antibody for red laser detection by flow cytometry

The Bad

None

The Bottom Line

For better results always use the antibody with eBioscience intracellular fixation & permeabilization buffer.

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