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Various Nucleofector™ Kits and corresponding Amaxa™ Optimized Protocols are available for the transfection of human CD34+ cells using the different Nucleofection™ Platforms. Optimal kits for transfection of human CD34+ cells in the 4D-Nucleofector™ and 96-well Shuttle™ System are the P3 Primary Cell Kits used in combination with cell-type specific protocols. Human CD34+ cell specific kits are available for the Nucleofector™ II/2b Device.
Example for Nucleofection™ of CD34+ hematopoietic progenitor cells with H-2Kk cDNA
Unstimulated peripheral blood mononuclear cells, partially enriched for CD34+ cells were transfected by Nucleofection™ using a plasmid encoding the mouse MHC class I heavy chain molecule H-2Kk. 16 hours post Nucleofection™ the cells were stained for CD34 and H-2Kk and analyzed by flow cytometry. Lymphocytes were gated according to forward/side scatter. Dead cells and CD34- cells were excluded by propidium iodide staining and gating. H-2Kk expression is shown after Nucleofection™ without (D) and with plasmid DNA (E).
For the 4D-Nucleofector™ Kits "L" and "S" indicate the Nucleocuvette™ Vessel format. L = 100 µl single Nucleocuvette™, S = 16-well Nucleocuvette™ Strip.