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Transient Transfection


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4 replies to this topic

#1 sorcener

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Posted 11 March 2010 - 03:32 PM

Hi! I hope somebody can help me,

I will make some probes overexpressing a protein but i'm not quite sure if transient transfections will give me enought time to measure what i want. Do somebody know how long plasmid remains on transfected cells? or how long the transfected protein expression can last?

many thanks!

#2 perneseblue

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Posted 11 March 2010 - 06:24 PM

Hi! I hope somebody can help me,

I will make some probes overexpressing a protein but i'm not quite sure if transient transfections will give me enought time to measure what i want. Do somebody know how long plasmid remains on transfected cells? or how long the transfected protein expression can last?

many thanks!


I don't know for sure. But it can last for about 2-4 days. It all depends on how much DNA was initially taken up by the cell (which is dependent on plasmid size, transformation method, cell type, amount of DNA used), cell line (ie length of cell cycle), and the protein being expressed.
May your PCR products be long, your protocols short and your boss on holiday

#3 Dukey

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Posted 13 March 2010 - 11:28 AM

Hi! I hope somebody can help me,

I will make some probes overexpressing a protein but i'm not quite sure if transient transfections will give me enought time to measure what i want. Do somebody know how long plasmid remains on transfected cells? or how long the transfected protein expression can last?

many thanks!


I don't know for sure. But it can last for about 2-4 days. It all depends on how much DNA was initially taken up by the cell (which is dependent on plasmid size, transformation method, cell type, amount of DNA used), cell line (ie length of cell cycle), and the protein being expressed.


I agree, probably 96 hours at most. You could make stable cells to get around this.

#4 DocFlow

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Posted 16 March 2010 - 06:42 AM

We transiently transfected 293E cells and harvested proteins for 3-4 weeks:

http://www.ncbi.nlm....pubmed/15847800

#5 sorcener

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Posted 19 March 2010 - 01:20 PM

[quote name='DocFlow' date='Mar 16 2010, 08:42 AM' post='62836']
We transiently transfected 293E cells and harvested proteins for 3-4 weeks:

Many thanks




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