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interest protein and total protein ratio range

293T protein ratio

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#1 mawson

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Posted 24 June 2012 - 06:10 PM

I would expresse a foreign protein in 293T by transient transfection. I want to know how may the weight rotio range of the foreign protein/total protein be. I failed to find some answers to this question,I wonder if someone would share some experience no matter about what kind of cells. thank you.

#2 bob1

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Posted 25 June 2012 - 01:10 PM

It all depends on how good the transfection is and how strong the promoter is for expressing the protein off the plasmid. I would expect that the ratio would be in the range of 1:1,000,000 or lower - often you can't even see a change in band intensity (of the correct size) on a coomassie or ponceau stained gel/membrane following SDS-PAGE of transfected cells.

#3 mawson

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Posted 25 June 2012 - 05:49 PM

It all depends on how good the transfection is and how strong the promoter is for expressing the protein off the plasmid. I would expect that the ratio would be in the range of 1:1,000,000 or lower - often you can't even see a change in band intensity (of the correct size) on a coomassie or ponceau stained gel/membrane following SDS-PAGE of transfected cells.

thank you for help. I used CMV promotor. As you said, I had tried SDS-PAGE and coomassie stain before, but there's no difference between the experiment and control. I supposed the raotio maybe 1:100, and this suppose with out proof maybe so far from the truth.




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