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