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ExGen 500 in vitro Transfection Reagent

#R0511	1 ml (for 100-200 in vitro transfections)
Related Documents (in pdf, ~30-50 KB): Certificate of Analysis: #R0511 MSDS (English) MSDS (English-USA) MSDS (German)

Description
ExGen 500 is a sterile solution of linear polyethylenimine (PEI) molecules (22 kDa) in water at a concentration of 5.47 mM in terms of nitrogen residues. PEI has a high cationic-charge density potential which makes it an excellent DNA condensing and gene-delivering agent (1, 2). Every third atom of the ExGen 500 backbone is a protonable amino nitrogen atom, which makes the polymeric association an effective “proton sponge” (1, 3). This property allows for endosome buffering and protects DNA from lysosomal degradation.
ExGen 500 is suitable for the transfection of a wide number of cell types (see the list) with high efficiency and minimal toxicity. Transfection can be performed in the presence or absence of serum. ExGen 500 showed superior results when compared to many other lipid or polymer based transfection reagents (figure 1).
Comparison of ExGen 500 with branched PEI (PEI 800 kDa, and PEI 25 kDa) showed that:

• ExGen 500 is more efficient in its ability to compact (4) and to transfect small quantities of DNA.

• ExGen 500 transfection efficiency is not affected by the presence of serum (4, 5).

Concentration
The solution is 5.47 mM in terms of nitrogen residues.

Reagent not Supplied
0.15 M NaCl (endotoxin-free, sterile) solution for dilution of DNA.

DNA Quality Requirements
The A₂₆₀/A₂₈₀ ratio should be at least 1.8. It is recommended to have DNA preparation containing the minimal amount of contaminant endotoxins.

Cells Successfully Transfected with ExGen 500

Considerations for Transfection with ExGen 500

Figure 1. 293 cells (Human Kidney Epithelioma) were transfected according to manufacturer's protocols
A. ExGen 500	B. Competitor's liposome based reagent
Figure 2. ExGen 500 used to express beta-galactosidase (A, B, C) and EGFP (D) in rat sympathetic neurons
Figure 3. C0-transfection of perinatal rat SCG neurons with beta-galactosidase (B) and EGFP (C) genes using ExGen 500
Horbinski, C., Stachowiak, M.K., Higgins, D., Finnegan, S.G., Polyethyleneimine-mediated transfection of cultured postmitotic neurons from rat sympathetic ganglia and adult human retina., BMC Neuroscience 2001, 2:2, published by BioMed Central Ltd. http://www.biomedcentral.com/1471-2202/2/2

References

1. Godbey, W.T., Wu, K.K., Mikos, A.G., Poly(ethylenimine) and its role in gene delivery, J. Controlled Release, 60, 149-160, 1999.

2. Goula, D., et al., Size, diffusibility and transfection performance of linear PEI/DNA complexes in the mouse central nervous system, Gene Therapy, 5, 712-717, 1998.
3. Behr, J.P., L'eponge a protons: un moyen d'entrer dans une cellule auquel les virus n'ont pas pense, Medicine/Sciences, 12, 56-59, 1996.
4. Boussif, O., Thêse de doctorat, ULP Strasbourg, Decembre 1996.
5. Ouatas, T., et al., T3-dependent physiological regulation of transcription in the Xenopus tapdole brain studied by polyethylenimine based in vivo gene transfer, Int. J. Dev Biol., 42, 1159-1164, 1998.

Updated gegužės 29, 2008 10:25