The invention provides an analysis method of pulp proteome in a cucumis melo.L. The method comprises the following steps: extracting pulp protein in the cucumis melo.L, firstly performing isoelectric focusing electrophoresis, and then performing SDS-PAGE gel vertical electrophoresis. The invention provides an economic, simple, easily-implemented and fast method suitable for the extraction and dimensional electrophoresis of total protein in the pulp of the cucumis melo.L. The protein is extracted through a Tris/phenol extraction-methanol/ammonium acetate precipitation method, the protein in the pulp of the cucumis melo.L can be effectively extracted, and the influences from mass sugar and other organic matters are reduced; at present, the repeated experimental results show that the experimental sample is comprehensive in preparation, the 2-DE electrophoresis operation is simple, the graph is clear, the repeatability is good, the reagent is low in toxicity and the result is reliable, a technical example is provided for the research of proteomes of the cucurbitaceae horticultural plants, and the method is suitable for analysis of the pulp proteome in the cucumis melo.L by the dimensional electrophoresis technology.