CD14 AF00 for Monocytes Separation

Immunology
The Jackson Laboratory for Genomic Medicine
Lab Manager, RA III

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Alexa Fluor700 anti-human CD14 Antibody

Catalog Number:

325614

This antibody was used in a pediatric study involving healthy and asthmatic patients. Multiple phenotyping panels were used to screen these patients for any changes in cell populations. Monocytes are part of one of these panels. This antibody works well to separate the classical monocytes CD14+ CD16- from intermediate monocytes CD14+ CD16+.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMC/monocytes from whole blood

Primary Incubation

2/100 dilution in final volume of 50ul (PBS+ 2%FBS) for 15min at RT at dark.

Blocking Agent

FcR block (Miltenyi) for 3min at RT

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

LSRII (Becton Dickinson)

Results Summary

Nice separation of CD14+ monocytes from whole blood and easy separation of classical monocytes CD14+ from intermediate monocytes by adding CD16 antibody (BV570, cat#302035, Biolegend).

Additional Notes

Stock concentration of 0.5Mg/ml was diluted to a working concentration of 20ug/ml for a final concentration of 1ug/ml during staining.

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Summary

The Good

Nice separation

The Bad

None

The Bottom Line

Nice separation of CD14+ monocytes from whole blood.

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