Protocol- Efficient transfection in macrophage-derived cells and primary macrophages
- Easy-to-use protocol
- Compatible with serum and antibiotics
jetPEI™-Macrophage transfects macrophages and macrophage-like cells that are considered difficult to transfect. It contains a mannose conjugated linear polyethylenimine that binds to cells expressing mannose-specific membrane receptors, such as macrophages.
jetPEI™-Macrophage is also the reagent of choice for the transfection of cell lines such as RAW 264.7.
INTELLECTUAL PROPERTY:
The use of polyethylenimine (PEI) or polypropylenimine (PPI) or cationic polymers similar in structure thereto for transfecting cells, as well as compositions comprising these cationic polymers and at least one nucleic acid, are the subject matter of U.S. Patent No. 6,013,240, EP Patent No. 0770140 and foreign equivalents, for which Polyplus-transfection™ is the worldwide exclusive licensee.
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Complexes formation is perfectly performed with NaCl solution. 0.5 ml of jetPEI™-Macrophage is sufficient to perform 250 transfections in 24-well plates. |
As shown in Figure 1, jetPEI™-Macrophage allows successful transfection of macrophages derived from monocytes maturated for 7 days in presence of GM-CSF.
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Figure 1. Human macrophages expressing Beta-Galactosidase after transfection using jetPEI™-Macrophage. |
Primary human macrophages and murine RAW 264.7 cells express significantly higher protein levels when transfected with jetPEI™-Macrophage (Fig. 2).
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Figure 2. Comparative efficiency of jetPEI™-Macrophage versus jetPEI™ on primary human macrophages and murine RAW 264.7 cells in 24-well plates. Primary human macrophages were transfected in the presence of 100 U/ml GM-CSF and 10% serum, using 1 µg pCMVLuc and 2 µl transfection reagent. Murine RAW 264.7 cells were transfected using 2 µg pCMVLuc and 6.4 µl transfection reagent. Luciferase activity was measured 24 h post-transfection. |
The jetPEI™-Macrophage protocol is as simple as that of jetPEI™: Mix the DNA with the reagent to form complexes and simply add the mixture to the cells. jetPEI™-Macrophage is compatible with serum and antibiotics, thus eliminating the need for media changes. Protein expression is determined 24 h to 72 h post-transfection.
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2. Condon C., S. C. Watkins, C. M. Celluzzi, K. Thompson and L. D. Falo (1996) DNA-based immunization by in vivo transfection of dendritic cells. Nature Med 2, 1122-1128
3. Erbacher P., M. T. Bousser, J. Raimond, M. Monsigny, P. Midoux and A. C. Roche (1996) Gene transfer by DNA/glycosylated polylysine complexes into human blood monocyte-derived macrophages. Hum Gene Ther 7, 721-729
