Description
GenJet™ DNA In Vitro
Tranfection Reagent for MCF-7 is pre-optimized for transfecting
MCF-7 cells. MCF-7 is a breast cancer cell line was isolated in 1970 from
a 69-year-old Caucasian woman. This cell line retained several
characteristics of differentiated mammary epithelium including the ability to
process estradiol via cytoplasmic estrogen receptors and the capability of
forming domes. This cell line expresses the oncogene Wnt7B. Tumor necrosis
factor alpha (TNF alpha) inhibits the growth of MCF-7 breast cancer cells.
Treatment with anti-estrogens can modulate the secretion of insulin-like growth
factor binding protein’s.
Refer to the following optimal transfection conditions for maximal transfection
efficiency on MCF-7 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to
600 transfections in 24 well plates or 50 to 100 transfections in 6 well plates.
Summary of Optimal Transfection Conditions:
Confluence on the day of transfection
|
90% |
| Cell culture conditions |
DMEM with 4.5 g/L glucose, 10% FBS |
| GenJet™ (µl) : DNA (µg) Ratio |
3:1 |
Diluent for DNA and Transfection Reagent
|
Serum-free DMEM with 4.5 g/L glucose
|
| Incubation Time to Form GenJet™/DNA Complex |
15 minutes at RT |
| Presence of Serum/Antibiotics during Transfection |
Yes |
| Change Medium After Transfection |
No |
| Maximal Efficiency |
36 hours |
Transfection Results:
Reporter Gene
|
EGFP |
Plasmid
|
pEGFP-N3 (CMV promoter)
|
| Efficiency (GFP %) |
75% |
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
GenJet™ reagent is
optimized for MCF-7
cells. MCF-7 A2 cells were grown per ATCC recommended culture medium and
transfected with pEGFP-N3 by GenJet™. The efficiency was checked 36 hours post
transfection
Data Sheet 
