Indirect Flowcytometry - (Jun/21/2011 )

M13 (alone) binds to the target cells BUt not in a specific manner, because it doesn't have any scFv on its surface. On the other hand, I am not getting too high background, about 15 to 20%. BUT the problem is that my test conditions show the same amount of fluorescence or less.

then I guess something is not working,
if I were you, I will fix the sample (target cell) before staining.
capping might be the reason you are not getting signal.

if you could have some kind of positve ctrl, that would be nice though.
or you could try immunocytochemistry to see whether you can see the signal under fluorescent microscope.

Hi again after apx 9 months!!!
I've finished my thesis and defensed it n now that I m trying to write my article, i do not know how to discuss the negative result of my flowcytometry...
Hope u be fine n sound