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I am using eBiosciense magnetic beads kit for MACS separation of murine CD11b+ cells (= monocytes and granulocytes).

It consists of Biotin-labelled anti-CD11b Ab; and a secondary agent: streptavidin crosslinked to magnetic particle.

But when afterwards running flow cytometry, with fluorochrome-labelled anti-CD11b Ab, I'm getting quite poor CD11b+ enrichment yield.

Does it mean that MACS did not work properly?

Or, maybe, that Biotin-linked anti-CD11b Ab which is used for MACS, which has already stuck to antigen, obstructs fluorochrome-linked Ab and prevents it from contact with Ag?

After MACS separation, any ways exist to wash off MACS reagents off the cells surface?

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