Description
GenJet™ DNA In Vitro
Transfection 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.
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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
