In vitro hepatocytes transfection

With Targefect-Hepatocyte (HEP-01) from Targeting Systems

The kit comes with two tubes: Targefect-Hepatocyte and virofect.

Before starting the experiment, thaw the reagents and vortex Targefect-Hepatocyte at full speed for 30 seconds, just before preparing the transfection complexes

  1. Grow hepatocytes in 5ml plates (DEME+5%FCS+P/S).
  2. Aliquot DNA. For 5ml plates, use 8 ug DNA for each plate.
  3. For each plate, mix DNA with 1.2 ml medium (hepatocytes medium) and flick for 10 seconds to make them mix well.
  4. Add 16 ul Targefect-Hepatocyte solution to the medium with DNA and flick for 10 seconds.
  5. Add 32 ul virofect solution to the medium and mix well.
  6. Incubate at RT in hood for 20 min.
  7. Suck the medium off in the plates.
  8. Add the transfection medium in and incubate for 2 hr in the incubator.
  9. Add 4 ml medium.

Culture Dish Medium in the mix DNA Targefect-Hepatocyte virofect Adding medium (2hrs later)
12 well plate 0.3ml 2ug 4ul 8ul 0.8ml
6 well plate 0.6ml 4ug 8ul 16ul 1.4ml
5ml plate 1.2ml 8ug 16ul 32ul 4ml