Transient transfection of primary cultured hepatocytes using CaPO4/DNA precipitation
We present a detailed protocol for the transient transfection of non-proliferating primary cultured hepatocytes that is easily reproducible. Using a modification of the classical CaPO4/DNA precipitation method, this protocol is an inexpensive alternative to other methods that are often cumbersome, e...
| Veröffentlicht in: | BioTechniques. - 1988. - 20(1996), 5 vom: 01. Mai, Seite 826-30, 832 |
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| Weitere Verfasser: | , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
1996
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Journal Article Calcium Phosphates alpha-tricalcium phosphate tetracalcium phosphate calcium phosphate, monobasic, anhydrous 701EKV9RMN DNA 9007-49-2 calcium phosphate 97Z1WI3NDX mehr... |
| Zusammenfassung: | We present a detailed protocol for the transient transfection of non-proliferating primary cultured hepatocytes that is easily reproducible. Using a modification of the classical CaPO4/DNA precipitation method, this protocol is an inexpensive alternative to other methods that are often cumbersome, expensive, difficult to reproduce or harmful to primary hepatocytes. Because only 0.5 x 10(6) cells are needed for a single transfection experiment, several reporter genes can be introduced into hepatocytes of a single liver preparation. With our protocol, different plasmids can be introduced into one cell. In this way, cis-trans interactions can be examined and reporter gene expression can be normalized for transfection efficiency. Furthermore, we describe details of a transfection experiment with two different reporter gene vectors using a luciferase gene and a lacZ gene. The results presented may be helpful to other groups concerned with improved timing of transfection experiments |
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| Beschreibung: | Date Completed 26.09.1996 Date Revised 28.09.2018 published: Print Citation Status MEDLINE |
| ISSN: | 1940-9818 |