The invention discloses a western blot detection method for phosphoprotein. The western blot detection method for the phosphoprotein comprises the following steps: S1, preparing a sample, carrying outgel electrophoresis, namely adding 6 times SDS sample loading buffer solution in a ratio of 5:1 into the phosphoprotein sample, uniformly mixing, and carrying out electrophoresis at a voltage of 80 V; S2, transferring a membrane, namely connecting a gel surface with a negative pole, connecting a PVDF membrane with a positive pole, and carrying out transfer printing for 1 hour at a constant voltage of 130 V; S3, sealing, namely putting the PVDF membrane into PBST containing 5% of BSA, and hatching for 20 minutes in a shaking table and at room temperature; S4, adding primary antibodies, namelyadding the primary antibodies, hatching for 2 hours in the shaking table and at the temperature of 24 DEG C, and rinsing for four times with PBST, wherein the rinsing is carried out once every 5 minutes; S5, adding secondary antibodies, namely adding anti-rabbit secondary antibodies labelled by horse radish peroxidase (HRP), hatching for 1 hour in the shaking table and at room temperature, and rinsing for four times with PBST, wherein the rinsing is carried out once every 5 minutes; and S6, developing, namely uniformly dropwise adding a developing reagent on the PVDF membrane, and carrying outgel developing. The western blot detection method for the phosphoprotein has the advantages that a wet membrane transferring manner is adopted during western blot detection of the phosphoprotein, anda protein signal is continuous and high in strength; and no non-specific band exists when dilution ratio of the primary antibodies is 1:5000, and the protein signal is better in effect.