This is not to say that private charities should not exist. They absolutely should, and participation in civil society is an important part of an individualâs and a nationâs ability to thrive. But a robust civil society does not require a separate ânonprofitâ sector.
Hi Athanassios. Remove media, wash cells with PBS 2-3 times, fix with 3.7% paraformaldehyde for 10 minutes, and Wash the fixed cells 2-3 times with PBS. After washing, permeabilize the cell monolayer by adding 0.1% Triton X-100 in PBS and incubate for 15 minutes at 37 CÂ°.
Transfecting HEK 293 Cells. Use this procedure to transfect plasmid DNA into HEK 293 cells in a 24-well format (for other formats, see Scaling Up or Down Transfections, below). All amounts and volumes are given on a per well basis. The day before transfection, trypsinize and count the cells. Plate 0.5 -1.25x105 cells per well in 0.5 ml of complete growth medium. Cell density should be 50-80% confluent on the day of transfection. (Optional) The day of transfection, remove growth medium from cells and replace with 0.5 ml of complete growth medium.