UCSF

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 (2hr 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