( were determined by measuring ...)Effectene Transfection Reagent

were determined by measuring luciferase activity 48 h post-transfection, and are given as relative light units (RLU). (Data kindly provided by I. Clavereau, D. Petitprez, and I. Van Seuningen, Unit INSERM 377, Place de Verdun, Lille Cedex, France.)


Transfection of Oligonucleotides Using Effectene Reagent



C6 glioblastoma cells (1 x 104) were transfected with either naked FITC-labeled CD44 antisense oligodeoxynucleotide (ODN) or with EffecteneODN complexes. Intracellular accumulation and distribution of the transfected ODNs was analyzed 16 h post-transfection by immunofluoresence. A Naked ODNs (2 g). B ODNs (0.04 g) complexed with Effectene Reagent. Transfected ODNs are green in the photos. (Data kindly provided by G. Beutel and M. Schott, Institute of Neuropathology, Hanover Medical School, Hanover, Germany.)

Procedure

The Effectene procedure has two steps. DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 25 minutes. Effectene Reagent is then added and the mixture is incubated for 510 minutes to allow EffecteneDNA complexes to form. The complexes are mixed with growth medium (which can contain serum and antibiotics), and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression.

Applications

Effectene Reagent is suitable for transient and stable transfection of a broad rang
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