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Efficient transfer of antibodies into mammalian cells by electroporation
J Lukas, J Bartek, M Strauss
Language English Country Netherlands
Grant support
IZ95
MZ0
CEP Register
Digital library NLK
Full text - Část
Source
NLK
ScienceDirect (archiv)
from 1993-01-01 to 2009-12-31
PubMed
8158003
Knihovny.cz E-resources
- MeSH
- Cell Line MeSH
- Cyclin D1 MeSH
- Cyclins immunology MeSH
- Electroporation * MeSH
- Fluorescent Antibody Technique MeSH
- Haplorhini MeSH
- Immunoglobulin G immunology MeSH
- Binding, Competitive MeSH
- Humans MeSH
- Microinjections MeSH
- Antibodies, Monoclonal immunology metabolism MeSH
- Mice MeSH
- Tumor Cells, Cultured MeSH
- Breast Neoplasms MeSH
- Oncogene Proteins immunology MeSH
- S Phase immunology MeSH
- Cell Survival MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
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.
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- $a 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.
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