Anti-human CD19 Microbeads - A Simple Method to Obtain Highly Pure B Cells from PBMC

May 05, 2016

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

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Miltenyi

Product Name:

CD19 Microbeads, human

Catalog Number:

130-050-301

The primary goal of our research was to obtain highly pure CD19+ B cells, as well as other immune cell types, from human peripheral blood mononuclear cells. The cells were required to be highly pure since downstream analysis involved microarray analysis of gene expression. This product was selected over competing products in part due to its ease of use and the previously established protocols provided by the manufacturer.

Experimental Design and Results Summary

Application

Positive selection of CD19+ 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 according to well established protocols. CD19+ microbeads were used to isolate CD19+ cells from PBMC following the exact protocol defined in the product datasheet when using an autoMACS Pro Separator. Four different cell types were positively selected in sequential order from the same human PBMC sample, with CD19+ selection occurring first.

Tips

When purifying more than one cell type from a single PBMC sample, you may want to consider purifying B cells first using the CD19+ microbeads, since B cells can bind other beads non-specifically through FcR - antibody interactions. Alternatively, as suggested by Miltenyi, you may want to use FcR blocking reagent in subsequent positive selections as a preventative measure.

Results Summary

A highly pure fraction of CD19+ B cells was obtained using these microbeads for positive selection.

Additional Notes

None

Image Gallery

Summary

The Good

Simplicity of the protocol. Ease of use. Established protocols.

The Bad

Knowledge of flow cytometric procedures and protocols is required to perform extra validation steps after using CD19+ beads to isolate B cells.

The Bottom Line

If you want to obtain a highly pure population of CD19+ B cells from a heterogeneous population of cells such as PBMC, this product is highly recommended and should be seriously considered.

Join the discussion