Expression of Expression Of CD64 (FcgRI) On Peripheral Human Monocytes Isolated From Patients With A Rare Autoimmune Disease.

January 23, 2017

Surgery
George Washington University, Washington DC
Post-doctoral Fellow

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Company:

BioLegend

Product Name:

PE/Cy7 anti-human CD64 Antibody

Catalog Number:

305022

CD64, the high-affinity IgG receptor FcgRI, is an integral membrane glyccoprotein that binds to monomeric IgG and potentiates inflammatory responses. CD64 is expressed by monocytes, macrophages, dendritic cells and neutrophils. Expression of CD16 along with CD14 and CD64 are crucial to determine and distinguish between classical and non-classical monocytes. Expression of CD64 indicates M1 type monocyte/macrophage which mediates type 1 inflammation and its over-expression is often responsible for tissue destruction contributing to an auto-immune response.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC or whole blood

Primary Incubation

100ul of peripheral blood was stained with 1ul (1:100) of anti-CD64 PECy7antibody. 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 CD64 was accessed.

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow cytometric detection

Results Summary

There was a high expression of CD64 among peripheral monocytes as observed by the high intensity staining of PECy7 CD64 (Blue FL-4 channel). Peripheral blood monocytes from patients with this particular auto-immune disease had elevated CD64 expression on monocytes reflecting a MI phenotype mediating type 1 inflammation.

DOI or PMID #

NA

Additional Notes

Standardization of the concentration of antibody used is important. Generally a 1:100 dilution is a good starting point.

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Summary

The Good

Easy and reliable method for analysis of surface expression of CD64 (FcgRI receptor).

The Bad

Nothing to mention

The Bottom Line

PE/Cy7 anti-human CD64 Antibody from Biolegend is a reliable antibody for analysis of monocyte/ macrophage phenotypic expression and activation state from peripheral blood or isolated PBMCs.

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