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Various Nucleofector™ Kits and corresponding Amaxa™ Optimized Protocols are available for the transfection of human B cells using the different Nucleofection™ Platforms. Optimal kits for transfection of human B cells in the 4D-Nucleofector™, 96-well Shuttle™ and HT Nucleofector™ System are the P3 Primary Cell Kits used in combination with cell-type specific protocols. Human B cell specific kits are available for the Nucleofector™ II/2b Device.
Example for Nucleofection of CD19+ B cells with H-2Kk cDNA
Fresh, unstimulated PBMC were transfected using a plasmid encoding the mouse MHC class I heavy chain molecule H-2Kk. 16 hours post Nucleofection™, the cells were stained for CD19 and H-2Kk. Lymphocytes were gated according to forward/side scatter (A). Dead cells and CD19- cells were excluded by staining with propidium iodide and gating (B, C). H-2Kk expression is shown post Nucleofection™ without (D) and with plasmid DNA (E).