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目的:测定4品种家蚕蛾下颚溶茧酶含量,并建立斑点免疫印迹法在中药材药效成分含量的测定方法。方法:以兔抗溶茧酶多抗为一抗,应用斑点免疫印迹法对4品种家蚕蛾下颚溶茧酶含量作定性检测,杂交信号应用Quantity one软件作定量分析。结果:4品种家蚕蛾下颚的个体溶茧酶平均含量分别为6.41、8.09、7.62、1.88μg/头。结论:不同品种家蚕蛾间下颚溶茧酶含量差异显著。斑点免疫印迹法简便快捷,结果准确,无需大型设备,可用于类似中药材药效物质的含量测定与品质评价。
OBJECTIVE: To determine the content of chymotrypsin in four species of silkworm moth, and to establish a method for the determination of medicinal components of Chinese medicinal materials by dot blot immunoblot assay. METHODS: Rabbit polyclonal antibodies against chymotrypsin were used as primary antibody. The content of Enzymes in four species of B. mori was determined by dot blot immunoblotting. Quantitative analysis of hybridization signals was performed using Quantity one software. Results: The average contents of individual chymase enzymes in the four species of silkworm moths were 6.41, 8.09, 7.62, 1.88 μg/head. Conclusion: There is a significant difference in the content of enzymes between different species of silkworm moth. Dot immunoblotting is simple, rapid, accurate, and does not require large equipment. It can be used for the determination and quality evaluation of medicinal substances similar to Chinese herbal medicines.