A method for detecting protein kinase based on surface plasmon resonance comprises the following steps: A, a gold film is used as a sensing layer, the sensing layer, first, is soaked with ethanol, then ultrasonically cleaned, H2 flame is used for annealing on the surface of the gold film; B, the gold film processed by the step A is soaked in a polypeptide P1 solution with an amino acid sequence shown as SEQIDNO:1, then soaked with the gold film in a polypeptide P2 solution with an amino acid sequence shown as SEQIDNO:2, and blown to dry with N2, and sealed storage of the prepared polypeptide P1 and P2-modified sensing gold film is performed at-18 DEG C for standby use; and C, the polypeptide P1 and P2-modified sensing gold film prepared by the step B is put on a SPR (surface plasmon resonance) apparatus, protein kinase flow injection is performed on the sensing gold film, when the sensing gold film is bonded with polypeptides on the surface of the gold film, a new resonance angle is produced, and the size of the resonance angle is a SPR response signal.