MagVigen™ – anti-CD45 magnetic nanoparticles are ideal for rapid isolation or depletion of human CD45+ progenitor stem cells. MagVigen™ magnetic nanoparticles coated with anti-CD45 monoclonal antibody recognize and efficiently bind to CD45+ cells following a short incubation. The nanoparticle bound CD45+ cells can be separated from the rest of the sample by magnet.
MagVigen™- anti-CD45 offers high recovery of high-purity and viable cells for use in further downstream molecular assays. The beads bound cells can be lysed for further protein or nucleic acid analysis. MagVigen™ nanoparticles are much smaller than conventional micro-beads. This feature allows for better accessibility of the nanoparticles to the antigenic epitope on cell surfaces. In addition, the surfaces of MagVigen™ nanoparticles are uniquely coated to reduce non-specific interactions with non-targeted CD45 negative cells.
- MagVigen™ – anti-CD45 magnetic nanoparticles are provided in 1 ml of phosphate buffered saline (PBS), pH 7.4.
Species Reactivity: Human. Others not tested.
All materials should be stored at 4°C.
Shelf life: 6 months
Protocol: Deplete or positively isolate CD45+ cells
- Dilute blood with an equal volume of PBS +2mM EDTA.
- Slowly layer the diluted blood over the Ficoll-Hypaque solution in a 50-ml conical centrifuge tube. Use 2ml of Ficoll-Hypaque (not included, product by Sigma Aldrich) per 1ml of blood.
- Centrifuge at 400g for 40min at room temperature with no brake.
- Collect the mononuclear cells located at the interface between plasma and the Ficoll-Hypaque and transfer to 15-ml conical tube.
- Dilute aspirated mononuclear cells with 4 volume of cold PBS.
- Centrifuge at 400g at 4°C for 10min and discard the supernatant.
- Re-suspend mononuclear cells from 1ml peripheral blood in 1ml of Ca2+ and Mg2+ free PBS buffer supplemented with 0.05% BSA and 5 mM EDTA.
- Wash 25μl of MagVigen-CD45 beads twice with 200μl of PBS Buffer. Re-disperse in 25μl of PBS buffer.
- Add pre-washed beads to the mononuclear cell solution with 4-5 million cells and incubate with gentle rotation at 4°C for 1-2 hour.
- Place the tube by a magnet for 5 min.
- For depletion: Transfer supernatant to a new tube for further use and discard the beads.
- For positive isolation: While the tube is still in the magnet, carefully remove and discard the supernatant.