Cell Line Generation

Transfection of Cell Lines

  • We determine the optimal seeding density (70% confluence) of your cell line of interest, and optimal antibiotic concentration (>90% cell death after 10 days) prior to transfection

  • For transfection, a delivered DNA construct is added to a cell line of interest in combination with Lipofectamine 3000 (optimized with different amounts of DNA and Lipofectamine). We then culture transfected cells in a flask in the presence of antibiotics to select transfected cell pools.

  • Knockdown is validated by immunoblotting. Transfection efficiency is determined via flow cytometry by staining the cells with an antibody targeting the respective protein, either on the surface or intracellularly

  • The cell lines with the highest protein expression are chosen for subsequent experiments (after single cell cloning upon request)

Oncolines® - Cell Line Generation - Transaction of cell lines