TransPass™ HUVEC Transfection Reagent
- This product was discontinued on 01/01/2013
This product includes 2 x 0.6 ml TransPass-V (store at -20°C) and 0.6 ml HUVEC Reagent Component (store at 4°C).
The TransPass™ HUVEC Transfection Reagent is designed specifically for transfecting endothelial cells including HUVEC, HMVEC, human aortic endothelial cells, etc. with optimal transfection efficiency (Figure 1). TransPass HUVEC Transfection Reagent consists of two components: the HUVEC Reagent Component, a non-lipid cationic transfection reagent, and TransPass-V*, an Adenovirus-derived component.
The addition of TransPass-V significantly enhances lipid-mediated transfection efficiency in many cell lines and primary cells including endothelial or epithelial (1,2).
The combination with the HUVEC Reagent Component yields optimal plasmid DNA transfection efficiency in HUVEC and many endothelial cell lines (Figure 2).
* TransPass-V contains a replication-deficient Adenovirus preparation. Because of the nature of this component, it should not be used with cell lines that contain Adenovirus sequences such as HEK-293, to avoid complementation of the virus. Additionally, it is recommended that common laboratory biosafety used in standard Adenovirus work should be practiced. For more information see http://oba.od.nih.gov/oba/index.html.
TransPass HUVEC and TransPass V are proprietary formulations manufactured by Targeting Systems. Please direct all inquiries regarding reagent compositions to Targeting Systems.
Cell Lines Successfully Transfected:
- HUVEC cells (Human umbilical vein endothelial cells (70-90%)
- Dermal microvascular endothelial cells (40%)
- Lung microvascular endothelial cells (70-90%)
- Human aortic endothelial cells (50%)
- Bovine aortic endothelial cells (60-70%)
- Rat endothelial cells (60-70%)
- Use early passages of cells (up to the 6th passage) for transfections.
- Use non-collagen and non-gelatin coated tissue culture plates only (transfection of cells plated on collagen-coated surfaces may result in lower transfection efficiency).
- Recommended media: MCDB131 from VEC technologies, Media 199 plus supplements and 20% fetal calf serum or EBM from Cambrex plus supplements and 10% fetal calf serum. Cells grown in these media appear healthier and give higher transfection efficiency.
- Clonetics HUVEC cells have given very good transfection results.
- For consistent results, it is important to maintain healthy proliferating cells that are regularly passaged.
- If cells have been grown in medium containing heparin, they must be washed after trypsinizing and resuspended in growth medium without heparin and antibiotics/antimycotics before plating for transfection.
- It is important that NO heparin and NO antibiotics/antimycotics are present in the growth medium during transfection.
- Use sterile plasmid DNA purified by CsCl gradient centrifugation or column chromatography.
- The amount of plasmid DNA per transfection can be varied, and the ratio of HUVEC Reagent Component to TransPass-V should be kept between (1:1) and (1:2).
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