reporter gene -seap - expression of seap is very low (Jul/07/2007 )
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I've done promoter expression level analysis with SEAP, and results seemed fine to me... so you make me in trouble.
I think like genehunter that 5% should be ok and that P/S doesn't change things too much.
P/S should not affect SEAP expression. Normally, you should use the same culture medium for maintianing these cells post transfection. Your concern with residual AKP activity in the serum is valid and I think 5% FBS is fine for many cells.
One note: SEAP has long halflife and can accumulate over time during your experiment, therefore it does not serve the purpose as a reporter for promoter strength, because you should see your SEAP activity goes up over time. This may create some confusion under some setting. Reporter with short halflife, such as luciferase does not have this problem.
-genehunter-1-
QUOTE (genehunter-1)
SEAP has long halflife and can accumulate over time during your experiment, therefore it does not serve the purpose as a reporter for promoter strength
do you have a reference for that point? i would be particulary interested in.I've done promoter expression level analysis with SEAP, and results seemed fine to me... so you make me in trouble.
QUOTE (jhilmil)
with and without P/S in medium??? what FBS (final) conc u had used
i didn't do with and without P/S... and i use 10% serum.I think like genehunter that 5% should be ok and that P/S doesn't change things too much.
-fred_33-
fred_33: www.pnas.org/cgi/content/full/0606398104/DC1
-genehunter-1-
thank you
-fred_33-
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