TransPass™ HUVEC Transfection Reagent: Transfection Protocol

Overview

Use Table 1 to select the reagent volumes for various plate sizes.

Protocol

1. 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).
2. Mix plasmid DNA with serum-free DMEM (Table 1).
3. Add the appropriate volume of HUVEC Reagent Component.
4. Add the appropriate volume of TransPass V.
5. Gently mix the transfection complex mixture by flicking the tube.
6. Incubate at room temperature for 20-30 minutes.
7. 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.
8. 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.