Fast, convenient, and effective method to transiently transfect primary hippocampal neurons.
Transfection of primary neurons in culture has proven to be experimentally challenging in the past. To overcome these limitations, we present a detailed transfection protocol for hippocampal neurons based on DNA/Ca(2+)-phosphate coprecipitation. The main advantages being (1) the speed and convenience, (2) the remarkable efficiency of transfection for mature neurons, and (3) consistent health of the neurons upon transfection allowing subsequent manipulations. The strength of this protocol is convincingly demonstrated by the fact that the expressed protein can be detected biochemically on Western blots.
Köhrmann
M
M
European Molecular Biology Laboratory, Cell Biology Programme, Heidelberg, Germany.
Haubensak
W
W
Hemraj
I
I
Kaether
C
C
Lessmann
V J
VJ
Kiebler
M A
MA
D000818:Animals; D018929:Cell Culture Techniques; D049452:Green Fluorescent Proteins; D006624:Hippocampus / Q000166:cytology* / Q000502:physiology*; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D051381:Rats; D014162:Transfection
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins