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[Bomber]

Hi everybody,

I have a question concerning transfection procedures:
I transiently transfected Raji cells by electropration and took a few samples for analysis in a western blot.
Surprisingly I noticed a strong decrease of a certain protein in comparison to non-transfected cells which I analysed on a western blot as well (the actin signals are equal).
I don´t really have an explanation for it: does anybody have an idea if this might be due to electroporation side effects, or are their any papers describing such things?

The vector codes for a shRNA: but the decrease of the protein can be detected as well as in my transfected control cells so I don´t think the effects are due to the used plasmids.

Thanks very much everybody and kind regards

[Scott]

Hi Bomber,

It is not surprising that after electroporation your cells will alter expression of some genes.  Passing a large electric shock through them will result in a number of physiological changes.  I haven't seen any papers on it, but it is something that must be remembered that a true control is not untransfected it is mock transfected cells.

Sorry I can't be more help

Scott