In my Nucleofection® Experiment, how critical is the time point for analysis post transfection?


It is very critical. Waiting too long for analysis post transfection can lead to loss of peak expression.

The optimal time for analysis is related to two factors: stability of protein being expressed and transfection method. For a short-lived protein (like luciferase) analysis should be done at 6-18 hours after Nucleofection®. For a more stable protein, such as GFP, one can wait to start analysis for up to 24 hours or longer post Nucleofection®. As Nucleofection® delivers DNA directly to nucleus at the time the pulse is delivered, expression will start much earlier than with other transfection methods. The best practice is to always perform a time point analysis when expressing a new protein. Further information on this topic you can find in the attached technical reference guide: "Important Vector Factors for Gene Expression".

Research Areas:
Gene Expression