Description
GenJet™ DNA In Vitro Tranfection Reagent for SHEP is pre-optimized for transfecting SHEP cells. This human neuroblastoma cell line was cloned from a single primary tumor. Unlike other neuroblastoma cell, SHEP cells does not produce IGF-II, expresses five-fold fewer IGF-IRs than SH-SY5Y and dies in serum-free media or following exposure to metabolic stressors.
Refer to the following optimal transfection conditions for maximal transfection efficiency on SHEP 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
|
85% |
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 |
Yes, ~5 Hours After Transfection |
Maximal Efficiency |
48 hours |
Transfection Results:
Reporter Gene
|
EGFP |
Plasmid
|
pEGFP-N3 (CMV promoter)
|
Efficiency (GFP %) |
80% |
Storage Condition
Store at 4 °C. If stored properly, the product is stable for 12 months or longer
Data Sheet