Anti-Human CD14 Microbeads – Quickly Obtain Highly Pure CD14+ Monocytes/Macrophages from PBMC

May 13, 2016

Memorial University of Newfoundland
Division of BioMedical Sciences, Faculty of Medicine
Research Assistant

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Quality of Results

Ease-of-Optimization

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Company:

Miltenyi

Product Name:

CD14 Microbeads, human

Catalog Number:

130-050-201

Our lab requires highly pure immune cell subsets, such as monocytes and macrophages, for our experiments. To purify monocytes and macrophages from human peripheral blood mononuclear cells (PBMC), we regularly employ Miltenyi anti-human CD14 microbeads for magnetic positive selection. This product was initially chosen because of the minimal optimization and straightforwardness of the protocol.

Experimental Design and Results Summary

Application

Positive selection of CD14+ cells from human PBMC using an autoMACS Pro Separator

Starting Material

Human Peripheral blood mononuclear cells (PBMC) isolated from whole blood

Protocol Overview

Human PBMCs were isolated from whole blood by standard Ficoll density gradient separation. CD14+ microbeads were used to isolate CD14+ cells from PBMC by positive selection according to the exact protocol outlined in the product datasheet when using an autoMACS Pro Separator.

Tips

Works best with freshly isolated PBMC; however, we routinely isolate from cryopreserved samples and obtain sufficient quantities of CD14+ cells for our downstream analysis. To maximize yield, we recommend you use this product on fresh, live cells.

Results Summary

A highly pure population of CD14+ cells was obtained. Yield was also excellent.

Additional Notes

None

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Summary

The Good

Detailed protocol comes with the product so that there is minimal optimization. Can be combined with other positive-selection microbeads so that multiple cell types can be obtained from a single biological sample if so desired. Purity and yield of isolated cells was excellent.

The Bad

Additional flow cytometric analysis required following separation for validation of the technique. Product is only available in one size.

The Bottom Line

If you want to isolate CD14+ monocytes or macrophages from human PBMC, this product should be seriously considered based on its ease of use and consistent performance.

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