Eletrophoresis - Ab caspasis 3 staining (Jan/13/2005 )

I have been working with rat C6 glioma cell line. The antibody used in our lab do not detect the caspase 3 protein in my cells with 50ug protein, but detect other kind of sample, total brain rat for example. What could be interfering in my experiment?

Thank's

Thay P

did you use the same protocol to prepare the protein extraction of C6 cell line and the rat brain? is your C6 protein extract not degraded? don't extract the same period with your cell line and rat brain for your protein sample. and do you know whether your cell line does express caspase 3? or 50 ug protein is not enough to detect the expression of caspase 3 in your cell line because of its low expression level. is caspase 3 related to APO? usually the tumor cell is resistant to APO. did you induce APO in your cell line to detect caspase 3? good luck!