发信人: Eureka (尤里卡), 信区: Biology
标 题: Re: How long does it take for expression
发信站: Unknown Space - 未名空间 (Tue Mar 2 22:00:06 2004) WWW-POST

Maybe you can try different protocals by using only EGFP, and check the cells
at differnt time points ,you will be able to see the EGFP expression by
noticing the green cells under fluorenscent microscope, so that you are able
to tell the time course of plasmid expression in different protocals.

And different plasmids have differnt expression patterns, I would try to
harvest transfected cells at different time points and measure the protein
level.

usually people harvest transient transfected cells in 36-48 hours, because in
tansient tansfection protein level may go down after a while, but the peak
time of protein level differs depends on the method, plasmids ,cell line, and
blah blah.

good luck!

【 在 glider (大苹果) 的大作中提到: 】
: How long does it take for transfected gene to be expressed in mammalian
cells
: after transient transfection?
:
: In my lab, some people took 36-48 hrs, then do the assays, some people using
: 24 hours... Furthermore, does different method need different time of
: incubation?
:
: what is the difference between lipofectamine 2000 and calcium phosphate
: transfection in this aspect?
:
: It would be great if you can share your experience!
:
:


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