I'm using MACS CD19 microbead(human) to isolate B cells from PBMC. But there is a monocyte contamination. Is there any soltuion to avoid monocyte contamination? (adhesion of monocytes on plates for hours is not an option)
Thank you in advance for the answers
Monocytes might be sticking to your column and eluting with the B cells. You could try the following.
1. Avoid using column based separation. You can find B cell negative selection kits from other suppliers (Stemcell Technologies/BioLegend) that use a bead based (no column) method. The added advantage is that you are labelling the unwanted cells and so your B cells would be untouched (ideal when doing downstream functional assays).
2. If you want to stick with MACS, you could use CD14 microbeads to remove monocytes first. Then use the monocyte depleted PBMCs (flow-through) for B cell isolation with CD19 microbeads.
Fc block could help. Or first a negative macs for monocytes (cd14?)
You could try first depleting monocytes using cd14 microbeads and the do you selection for cd19 + b cells
Thank you. i am going to try the fc blocker, and then if its not working i will looking for CD14 microbeads.
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