GeneXPlus Transfection Reagent
ACS-4004 ™
ACS-4004 ™
This reagent affords high levels of gene expression in a variety of cell types and is suitable for both transient and stable transfection. GeneXPlus works effectively in large scale reactions, producing high protein yields. Since the reagent is composed of animal-origin free components and is serum compatible, there is no need for any culture medium change after transfection.
GeneXPlus Transfection Reagent has been optimized for use with HEK293T/17 SF suspension cells (ATCC ACS-4500) and HEKPlus SFM (ATCC ACS-4002) to reproducibly express a wide range of proteins at high levels.
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Guidelines for optimal plasmid DNA transfection
Reaction conditions should be optimized for each cell type to ensure successful transfections. However, the general suggestions below have been demonstrated to yield high efficiency transfections using GeneXPlus Transfection Reagent. Table 1 presents recommended starting conditions based on culture vessel size.
Culture vessel | 96-well plate | 24-well plate | 12-well plate | 6-well plate | 10-cm dish | T75 flask | 125-mL shaker flask |
Surface area (cm2) | 0.35 | 1.9 | 3.8 | 9.6 | 59 | 75 | N/A |
Complete Growth Medium (mL) | 0.092 | 0.5 | 1 | 2.5 | 15.5 | 19.7 | 20 |
Diluent (serum-free medium) (µL) | 9 | 50 | 100 | 250 | 1500 | 1900 | 2000 |
Amount of DNA (µg) | 0.1 | 0.5 | 1 | 2.5 | 15 | 19 | 20 |
Transfection reagent (µL) | 0.3 | 1.5 | 3 | 7.5 | 45 | 57 | 60 |
Transient plasmid DNA transfection protocol per well of a 6-well plate
Note: Adjust volumes for GeneXPlus Transfection Reagent, DNA and complete growth medium based on the surface area of the cell culture vessel as described in Table 1.
Cell Seeding
Note: For higher transfection efficiency, it is recommended that the cells are > 80% viable at the time of transfection.
Preparation of Transfection Reagent:DNA complex (prepare immediately before transfection)
Addition of Complexes to Cells
For generating stable cell transfectants: Passage cells 24 to 48 hours post-transfection in complete growth medium containing appropriate selection antibiotics, such as G418 or Hygromycin B. Maintain selection for 1 to 2 weeks to allow for selection of cells that have undergone stable integration of DNA.
Large-scale transfections: Table 1 recommends starting volumes for 20-mL transfections (125-mL shaker flask). The volumes listed can be directly scaled up in proportion to the culture volume, if larger volumes are desired.
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Yang H, et al. Effect of TAK1 on osteogenic differentiation of mesenchymal stem cells by regulating BMP-2 viaWnt/β-catenin and MAPK pathway. Organogenesis 14(1):36-45, 2018. PubMed: 29913119