斑点免疫印迹法快速测定4品种家蚕蛾下颚溶茧酶含量

Rapid Determination on Cocoonase Contents in Maxilla of Four Silkmoth Breeds by Dot Blotting

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 cocoonase content in maxilla of four silkmoth breeds of Bombyx mori and set up a dot blotting method to determine the content of the drug action components in Chinese medicines. Methods ： The contents of cocoonase in maxilia of four silkworm breed were determined by dot blotting with rabbit antibody of anti-cocoonase as the first antibody. Blotting signals were analyzed by software of Quantity one. Results： Contents of cocoonase in maxilla of four silkmoth breeds were 6.41,8.09,7.62, and 1.88 ug/moth, respectively. Conclusion： The difference among four silkmoth breeds was significant in the content of maxilla cocoonase. Dot blotting, as a convenient and accurate method could be used for determination of the contents of drug action components in Chinese crude drugs similar to silkworm.