Description: Vaxim Inc. GeneXPlusTM-1 is a formulation of a neutral lipid and a proprietary cationic lipid derived from our recent innovation of gene delivery technology, through a rational molecular design based on cell endosome PH balance and disruption mechanisms. It has excellent performance under wide transfection reagent range, highest efficiencies in diverse cell types, very minimized cytotoxicity, and user-friendly protocol. It is adaptable to suspension and adherent cells. It is suitable for transfections in vitro and in vivo, and great for both transit and stable transfection. One vial of GeneXPlusTM-1 (0.6 ml) is sufficient for 150 transfections at 24-well plate formate.
Contents: GeneXPlusTM-1 is supplied in steril liquid form.
Storage: Store at 4°C.
Stability: GeneXPlusTM-1 is stable for at least 1 year at 4°C.
Highest efficiencies in most cell types.
Excellent performance in presence and absence of serum.
Very minium cytotoxicity suit best for stable gene transfection.
User-friendly transfection protocols for all experimental format.
Tested in vivo mouse model without toxicity up to 3 ml/mice.
For adherent cells: Plate the cells in 24-well plates at a density of 5 - 8 x 104 cells / well in 0.5 ml of complete media without antibiotics per well and place plates in a 37°C, 5% CO2 humidified incubator. After 24 – 48 hours, the cells reach 60 – 80% confluence, begin preparations for transfection:
For suspension cells: Split the cells the day before transfection. On the day of transfection, harvest cells by centrifugation, remove the medium. Seed 5.5 – 8.5 x 105 cells in 0.5 ml of complete media without antibiotics per 24-well plate, begin preparations for transfection:
a. Dilute 0.8 ug of DNA in 20 ul of deionized H2O containing no serum, proteins or antibiotics. Mix and spin down the solution for a few seconds to remove drops from the top of the tube.
b. In a separated vial, dilute 3 - 5 ul of GeneXPlusTM-1 tansfection reagent with deionized H2O to total 20ul.
c. Add the transfection reagent to the DNA solution. Mix by pipetting up and down 6 times.
d. Let the solutions incubate for 15 minutes (20 – 25°C) to allow transfection-complex formation.
e. Add 60 ul of DMEM with no serum and no antibiotics to the DNA-lipid complex to final volume of 100 ul.
Add immediately the transfection complexes drop-wise to the appropriate well. Gently swirl the plate to ensure uniform distribution of the transfection complexes.
Put the cells back into the incubator.
Incubate the cells with the tranfection mixtures for 24 hours to 48 hours.
For transient transfections, assay cells for expression of the transfected gene.
For stable transfections, passage cells 1:4 to 1:8 into the appropriate selective medium 24 – 72 hours after transfection, replace with fresh complete medium with antibiotics.
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