Use Table 1 to select the reagent volumes for various plate sizes.
- Plate cells at an appropriate density so they will reach 70-80% confluence at the time of transfection (plating media should contain 10-20% serum).
- Mix plasmid DNA with serum-free DMEM (Table 1).
- Add the appropriate volume of HUVEC Reagent Component.
- Add the appropriate volume of TransPass V.
- Gently mix the transfection complex mixture by flicking the tube.
- Incubate at room temperature for 20-30 minutes.
- Add the transfection complex mixture to cells (Do not remove the serum-containing medium). Rock the plate gently in order to evenly disperse the transfection complex mixture.
- Return the plate to the incubator and incubate 24-48 hours before assay.
Note: The recommended ratio of HUVEC Reagent Component to TransPass V is between 1:1 and 1:2.