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to check protein expression - (Aug/31/2006 )

I've transfected a gene cloned with GFP in human fibroblasts cell line & i want to have stable cell lines.The vector which i've used is a mammalian vector.I want to know if by any method i can check the expression of my protein just after transfection so that i don't have to wait for stable cell lines to form.

-minnee-

You could try RT PCR and/or western.

-I love MSGs!-

You can have a look at the cells using a fluorescence mucroscope: GFP gives you a green signal that you should be able to see

-dnafactory-

u will grow ur transfected cells in antibiotic media for selection, so this should give some idea.

and as dnafactory suggested, GFP fluorescence can confirm this.

-scolix-

suggested methods are only suitable in most cases after several hours mostly at least 8h, then cells may begin to express your GFP-tagged protein; transfection efficiency before is only detectable if you have labeled your transfection DNA with e.g. a fluorescent marker; then you can directly follow with a fluorescence microscope transfection efficiency (expression efficiency is a different thing and detectable with FM for GFP or Western)

-The Bearer-