Bright Staining Of CD31 On Human PBMC By Flow Cytometry Using An Anti-Human CD31 Monoclonal Antibody (Clone WM59)

August 01, 2016

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:

BioLegend

Product Name:

Alexa Fluor® 647 anti-human CD31 Antibody

Catalog Number:

303111

Our lab routinely uses flow cytometry to measure the expression of various molecules on human peripheral blood mononuclear cells under various experimental conditions. The specific goal of this experiment was to measure CD31 expression on human PBMC and compare across disease states. This antibody was chosen because it is labelled with a fluorophore and has been quality tested for flow cytometry by the manufacturer.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC isolated from a healthy adult donor by Ficoll density gradient separation

Primary Incubation

Alexa Fluor 647 anti-human CD31 antibody (BioLegend 303111), incubated with approximately 1x10^6 cells for 1 hour on ice at 1:20 dilution (0.5ug 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 (Becton Dickinson)

Results Summary

CD31 was expressed on a significant proportion of human PBMC (black dots, black histogram). Staining was bright and CD31-expressing cells were easily distinguished from negative cells and cells labelled with a matched isotype control (red dots, red histogram). Furthermore, cells which did not express CD31 resembled the isotype control. The CD31 expression profile was almost identical to that provided by the manufacturer.

Additional Notes

None

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Summary

The Good

Directly conjugated with a fluorophore. Minimal optimization. Bright staining of cells.

The Bad

None

The Bottom Line

Excellent for measuring CD31 expression on human lymphocytes, monocytes, and granulocytes. Performed exactly as described by the manufacturer with no optimization required.

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