GenJet™ In Vitro DNA Transfection Reagent for Caco-2 Cell

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.

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Background

Tips for selecting diluents for GenJet™ DNA transfection reagents.
For efficient formation of transfection complexes, it is essential to choose an appropriate diluent to dilute the DNA transfection reagent and DNA. In addition to temperature and incubation time, the nature of the diluent is very important for the complete formation of the transfection complex, thus significantly affecting the DNA transfection efficiency. According to our validation data, the appropriate diluent is 50 times more efficient than the "wrong" diluent.

Follow the following technical tips to choose the proper diluent for GenJet™ DNA transfection reagents:
1. For the complete formation of transfection complex, the diluent must be serum-free and protein free.
2. Try to use a diluent that is closest to the composition of cell culture medium. If you are using DMEM (supplemented with serum and antibiotics) to grow HEK293 cells, the best diluent is serum-free and antibiotics-free DMEM. If you are using RPMI-1640 Medium (supplemented with serum and antibiotics) to grow Hela cells, the best diluent is serum/antibiotic-free RPMI-1640 medium.
3. Do not use media containing unknown components. For example, never use Opti-MEM™ to dilute GenJet™ DNA transfection reagents. According to our validation data, diluting our transfection reagents and DNA with Opti-MEM™ at least sacrificed 50% transfection efficiency on HEK293, Hela, CHO and NIH 3T3 cells. In addition, Opti-MEM™ may contains low level of serum, which would greatly compromise transfection efficiency.
4. Basically serum-free DMEM with high glucose is pretty good choice. It is very easily accessible and compatible with other cell growth mediums. If you have trouble with lower transfection efficiency, you may think about changing diluent to serum-free DMEM with high glucose. If you do not know how to choose a proper diluent, you can try serum-free DMEM with high glucose first which usually gives you very good transfection efficiency.