In this work, we have designed an electrochemiluminescence (ECL) reaction system used to indicate P53 protein from human lung cancer cells. And we optimized the working electrode firstly in this experiment. The electrochemiluminescence magnetic immunoassay is quantitative, reproducible, and requires only minimal sample processing. A single photon counter was used to heighten the ECL detection limit. The homologous immunoassay format consisted of a double antibody sandwich in which a biotinylated capture antibody, pre-bound to streptavidin-coated magnetic beads, was used to bind antigen from test samples. A detector antibody, labeled with ruthenium( II ) tris-bipyridal chelate, was added and, when bounded to the bead immunocomplex, light was generated in the presence of an excess of tripropylamine. The light was detected and measured by the ECL reaction system we designed. High-sensitive, rapid and reproducible detection of tumor marker P53 protein was actualize with this detection system. The results showed that the electrode with many holes on it can enhance the ECL efficiency, so the detection limit of the system can be improved to 1/1000 fmol/L free labeled Ru(bpy)32+. Ru(bpy)32 + TPA(tripropylmine) ECL reaction system was used to indicate P53 protein from human lung cancer cells. This method could have potential applications in early-stage tumor diagnosis and environmental inspection.