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
Cell culture conditions
GenJet™ (µl) : DNA (µg) Ratio
Diluent for DNA and Transfection Reagent
Incubation Time to Form GenJet™/DNA Complex
Presence of Serum/Antibiotics during Transfection
Change Medium After Transfection
Maximal Efficiency
Transfection Results:
Reporter Gene
Plasmid
Efficiency (GFP %)
|
90%
DMEM with 4.5 g/L glucose, 10% FBS
3:1
Serum-free DMEM with 4.5 g/L glucose
15 minutes at RT
Yes
No
36 hours
EGFP
pEGFP-N3 (CMV promoter)
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