I have a planned experiment that involves the following

1) MACs sorting of specific primary cells (single cell suspension) from an inducible Cre mouse which is tamoxifen naive.

2) Grow these cells Ex-vivo, and treat with 4-OHT (Will tag specific cells from RFP to GFP via an endogenous mT/mG reporter)

3) Sort cells which are tagged (will be a heterogeneous RFP/GFP population,)

4) Grow the sorted, 100% GFP+ cells in culture

For the second sorting step (Step 3), is it possible to use MACs sorting again via an anti-FITC antibody? Or will beads from the initial sorting stem make this impossible? I would like to utilize MACs and not FACs so that I have a higher number of cells to plate but was unsure if you could sort cells a second time or not.

Any input would be greatly appreciated!

Thank you for your time!

Kyle