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.pdf