Expression Of CD16 (FcgammaRIII) On Monocytes From Human Peripheral Blood In An Autoimmune Disease

January 09, 2017

Surgery
George Washington University, Washington DC
Post-doctoral Fellow

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Company:

BioLegend

Product Name:

PerCP/Cy5.5 anti-human CD16 Antibody

Catalog Number:

302028

CD16 is expressed in Monocytes/Macrophages, NK cells and neutrophils and is also known as the FCgammaRII receptor. They bing to Fc portion of IgG and activate Antibody-dependent cellular cytotoxicity. Expression of CD16 along with CD14 and CD64 are crucial to determine and distinguish between classical and non-classical monocytes. I our experiment , the aim was to analyze expression levels of CD16 on monocytes from peripheral blood of human sample with a particular autoimmune disease.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human peripheral blood

Primary Incubation

100ul of peripheral blood was stained with 1ul (1:100) of anti-CD16 percpCy5.5 antibody. Cells were kept for 30 mins. at room temperature. After incubation cells were washed twice with 1XPBS and re-suspended in 500ul of staining buffer. Cells were then analyzed by flow cytometry where RBCs were gated out utilizing a threshold voltage.Monocytes/macrophages were gated according to Fsc and Ssc and expression of CD16 was accessed.

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow cytometric detection

Results Summary

There was a high expression of CD16 among peripheral monocytes as observed by the high intensity staining of PercpCy5.5 CD16 (Blue FL-3 channel). Thus we could determine that the sample human blood had elevated CD16 expression on monocytes reflecting expansion of non-classical monocyte and perhaps causing an inflammatory response.

DOI or PMID #

NA

Additional Notes

Staining intensity can be adjusted by a serial dilution of the antibody. In our case, we found the intensity to be on the higher side using 1:100 antibody dilution.

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Summary

The Good

Easy and reliable method for analysis of surface expression of CD16

The Bad

Nothing to mention

The Bottom Line

The PerCP/Cy5.5 anti-human CD16 antibody works well on human peripheral blood for the analysis of surface expression of human CD16.

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