GenJet™ In Vitro DNA Transfection Reagent for B16-F10 Cells

GenJet™ DNA In Vitro Transfection Reagent for B16-F10 cell is pre-optimized for trans fecting B16-F10 cells. B16-F10 is a derived murine melanoma cell line, which is well-established model for the study of experimental cancer therapies. Melanomas express different tumor-associated antigens, which are potential targets for novel designs of therapeutic cancer vaccines. Melanoma-associated antigens include tyrosinase and tyrosine-related protein (TRP)-1/2 or the MAGE and BAGE class of antigens. Many immunotherapeutic protocols have been tested using the murine B16-F10 melanoma cell line that originated in the syngeneic C57BL/6 (H-2b) mouse strain.

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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.