transfection of PC12 cell line - (Apr/30/2007 )

hi, all.

Interested protein is endogeous in PC12.
I will treat siRNA of the protein in pc 12 and study its function.
Before this experiment, I done transfection that protein in PC12.

BUT....
the protein NOT overexpression.

why not this protein don't transfection??

help. ^^

I assume you are asking why it did not overexpress in PC-12 after transfection with extra copies of DNA? Two things that you may want to test: 1) transfect and afterwards detect this protein on some easy-to-transfect cells, such as 293, or cos-1 or cos-7. This will tell you if your construct is functional. 2) Do a co-transfection with a reporter to see if your transfection was working on PC12 cells, EGFP by flouresence microscopy or beta-Gal with X-Gal histochemistry staining are some options.