Description
GenJet™ DNA In Vitro
Transfection Reagent for Caco-2 Cell is pre-optimized for transfecting Caco-2
cells. The Caco-2
cell line is an immortalized line of heterogeneous human epithelial colorectal
adenocarcinoma cells, developed by the Sloan-Kettering Institute for Cancer
Research through research conducted by Dr. Jorgen Fogh. The Caco-2 cell
line is widely used with in vitro assays to predict the absorption rate of
candidate drug compounds across the intestinal epithelial cell barrier. The
assay requires that drug absorption rates be determined 21 days after Caco-2
cell seeding to allow for monolayer formation and cell differentiation.
Caco-2 cells may also refer to a cell monolayer absorption model.
Cell-based functional assays, such as the Caco-2 drug transport model for
assessing intestinal transport, are extremely valuable for screening lead
compounds in drug discovery.
Refer to the following optimal transfection conditions for maximal transfection
efficiency on Caco-2 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 5 Hours 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
Yes
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
A Picture Showing Transfection Efficiency of GenJet™ Reagent on Caco-2 cell with
GenJet™ reagent.
Caco-2ela cells
were grown per ATCC recommended culture medium and transfected with pEGFP-N3 by
GenJet™. The efficiency was checked 48 hours post transfection
Data Sheet
