Good Antibody Against Human CD8 For Flow Cytometry

March 29, 2017

Division of BioMedical Sciences, Faculty of Medicine
Memorial University of Newfoundland
Research Assistant

Overall

Quality of Results

Ease-of-Optimization

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Company:

Affymetrix eBioscience

Product Name:

Anti-Human CD8a APC

Catalog Number:

17-0086-42

One of the main areas of focus of our lab is lymphocyte biology, with a focus on T cell function and how it relates to disease. Our goal in using this product was to identify CD8+ cells in human peripheral blood mononuclear cells. This particular product was chosen because it is directly labelled with a fluorophore and ships pre-diluted ready to use.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human peripheral blood mononuclear cells isolated from whole blood by Ficoll gradient separation

Primary Incubation

Approximately 1x10^6 cells were incubated on ice for 1 hour in the dark with Anti-Human CD8a APC (17-0086-42; clone OKT8) at a 1:5 dilution in a total volume of 100ul PBS+2% FCS, 1mM EDTA, 0.1% sodium azide.

Blocking Agent

N/A

Secondary Incubation

N/A

Tertiary Incubation

N/A

Detection

FACSCalibur

Results Summary

This antibody was used to identify CD8+ T cells in human PBMC by flow cytometry. This antibody gave excellent results in FACS staining with clear separation of CD8- and CD8+ cells (open histogram). Cells stained with a matched isotype control and shown as the grey filled histogram. CD8- cells resembled isotype control cells as expected.

DOI or PMID #

N/A

Additional Notes

For analysis, lymphocytes were gated on according to their expected forward vs side scatter profile.

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Summary

The Good

Good separation between positive and negative cells. Directly labelled with a fluorophore. Already pre-diluted to optimal working concentration, thus minimal optimization required.

The Bad

None

The Bottom Line

I highly recommend this antibody for staining CD8 on human samples for flow cytometry. With minimal optimization required and a standard, quick staining protocol resulting in good separation of positive and negative events, you can't ask for much better than that!

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