Efficient transfer of antibodies into mammalian cells by electroporation.
An optimized protocol for electroporation is described which is suitable for introducing antibodies into mammalian cells. The method results in the uptake of detectable amounts of antibodies in 80% of the cells and in 40% large amounts are introduced. As an example, cell cycle activity (transition from the G1 to S phase) was inhibited by the introduction of monoclonal antibodies against G1-specific cyclin D1 into CV-1 and MCF7 cells. This specific antibody mediated inhibition of cellular function did not affect the viability of the cells since they recovered from the inhibition after some time. While approaching the efficiency of microinjection, the new protocol for electroporation of antibodies additionally permits treatment of the larger number of cells which are required for biochemical analyses.
Lukas
J
J
Danish Cancer Society, Division of Cancer Biology, Copenhagen.
Bartek
J
J
Strauss
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D001667:Binding, Competitive; D001943:Breast Neoplasms; D002460:Cell Line; D002470:Cell Survival; D019938:Cyclin D1; D016213:Cyclins / Q000276:immunology; D018274:Electroporation; D005455:Fluorescent Antibody Technique; D000882:Haplorhini; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008845:Microinjections; D015513:Oncogene Proteins / Q000276:immunology; D016196:S Phase / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D016213:Cyclins; D007074:Immunoglobulin G; D015513:Oncogene Proteins; D019938:Cyclin D1