CD14: Good Signal, but Relatively High Background!

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Pharmacology
University of Kentucky
Postdoc

Company:

BioLegend

Product Name:

PE anti-human CD14 Antibody

Catalog Number:

325606

CD14 is the LPS receptor, expressed at high levels on monocytes. When staining ficoll-paque isolated human PBMC, the monocytic gate is not always accurate enough to identify monocytes; hence it is useful to stain for CD14. We selected this product because it fits well with the other antibodies and fluorochromes used.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBLs

Primary Incubation

5 microlitre antibody for 30 minutes on ice (recommeded by BioLegend)

Blocking Agent

Human IgG 1ug/ml solution

Secondary Incubation

Not used

Tertiary Incubation

Not used

Detection

Flow cytometry

Results Summary

The monocytes stained very brightly with CD14; however, despite a 4-fold dilution (using a forth of the recommended antibody volume), we still found some background signal. Also, the signal on monocytes got weaker with higher antibody dilutions.

Additional Notes

Events shown are from a combined lymphocytes and monocytes gate.

Image Gallery

Summary

The Good

Strong signal

The Bad

High background

The Bottom Line

A good antibody, but the high background lets it down.

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