Posted 22 January 2013 - 07:47 AM
I have some problems with intracellular staining. Im fixating my cells with pfa, and permeabilizing them with saponin, and use an isotype control mouse IgG1 FITC, the same as the specific antibody. However the isotype control is so reactive and as the cells are stimulated the percentage of isotype positive cells increase by 30 %. But the specific antibody just increase with 5 %. and shows much less positive cells than the isotype control. This is so frustrating!
I also tried another pair of antibodies, rat IgG1 PE, but it shows the same tendency. Any advice?