Description
GenJet™ DNA In Vitro Tranfection Reagent for L929 cell is pre-optimized for transfecting L929 cells. L929 is a murine aneuploid fibrosarcoma cell line which is often used to assay TNF-alpha and TNF-beta. Treatment with TNF initiates apoptosis and subsequent cell death. The level of detection is on the order of 50 pg/mL. Another cell line which is approximately 5-10-fold more sensitive is WEHI-164. Some L929 subclones have been developed that allow detection of TNF at concentrations of approximately 30 pmol/L (approximately 500 fg/mL). Sublines that are resistant to TNF have been established also.
Refer to the following optimal transfection conditions for maximal transfection efficiency on L929 cells. GenJet™ reagent, 1.0 ml, is sufficient for 300 to 600 transfections in 24 well plates or 150 to 300 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 %)
|
~80%
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
Yes, ~5 Hours After Transfection
48 hours
EGFP
pEGFP-N3 (CMV promoter)
65%
|
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
Data Sheet