Human CD8 APC-Cy7 for FACS

May 25, 2017

Microbiology & Immunology
Columbia University
MD/PhD Student

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Company:

Tonbo Biosciences

Product Name:

Anti-Human CD8a (RPA-T8) APC-Cyanine7

Catalog Number:

25-0088

This antibody was used in a multi-color FACS analysis of human PBMCS to differentiate memory subsets of CD8+ T cells by transcription factor expression. This clone has been used extensively for identifying human CD8+ T cells. The fluorophore was specifically selected to preserve other brighter channels for difficult antigens.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Healthy human PBMCS

Primary Incubation

1:100; 20minutes; on ice in FACS buffer

Blocking Agent

2% FBS in dPBS; 2mM EDTA

Secondary Incubation

None

Tertiary Incubation

None

Detection

BD Fortessa

Results Summary

Healthy human PBMCs were thawed from liquid nitrogen and stained with a multi-color panel to differentiate memory CD8+ T cell subsets. Cells were stained for surface markers and subsequently fixed/permeablized with eBioscience Foxp3 transcription factor kit. After exclusion of dead cells, live CD8+ T cells were successfully separated from bulk CD3+ T cells and subsequently could be gated on for analysis of transcription factor expression. Fluorescence intensity was preserved despite aldehyde fixation.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Inexpensive; reasonably separation; allows for complex panels; fluorescence preserved despite aldehyde fixation

The Bad

None

The Bottom Line

While it does not provide the largest separation of CD8+/- possible, this antibody is more than sufficient for differentiating CD8+ T cells in peripheral blood samples and provides a very economical option for laboratories seeking to create complex panels for human T cell subsets.

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