empty vector control - (Feb/18/2008 )
hey guys, i am trying to show expression of a protein by transitently transfecting PC12 cells. i seen fluorescence in the cells but i realize i need an empty vector control. my dilemma is that i don't know how i would show that my empty vector was taken up by the cells since i do not have a marker for that. i can't use the GFP vector because my secnodary antibody is conjugated to cy2, which has very similar excitation and emission spectra as GFP. any help would be much appreciated.
Can you show it through antibiotic resistance?
well, i was thinking about showing it through antibiotic resistance but i was hoping to avoid doing stable cell line transfections with the vector containing the control DNA. would antibiotic resistance be enough to show that my cells took up the vector that otherwise, does not contain any visual markers such as GFP?
Many of the vector kits that I use contain an expression control containing LacZ- do you have something like that available to you? Then you could stain the cells to show that they have taken up the plasmid.
You could if the control vector has the antibiotic resistance gene. In this case, you will transfect cells and select them in media with antibiotics as you would for stabel cells except that this clone will not express anything.