二维电泳在人舌下和颌下唾液蛋白质混合物分析中的应用

Two-dimensional electrophoresis in the analysis of a mixture of human sublingual and submandibular salivary proteins.

作者信息

Mogi M, Hiraoka B Y, Fukasawa K, Harada M, Kage T, Chino T

出版信息

Arch Oral Biol. 1986;31(2):119-25. doi: 10.1016/0003-9969(86)90035-x.

Abstract

Complex protein mixtures of unstimulated human sublingual and submandibular saliva were fractionated by two-dimensional (2-D) gel electrophoresis, visualized by silver staining and then analysed by immunostaining. Specific proteins were identified by incubation with specific antibody and peroxidase-conjugated second antibody (Western blot). Electrophoresis and silver staining revealed over 50 protein components in 2 microliter of unconcentrated mixture. The Western-blot technique allowed detection of protein spots of plasma origin when an antibody against whole serum was used, but only the albumin spot could be found. Albumin, secretory IgA, acid phosphatase and alpha-amylase were identified with specific antibodies.

摘要

未刺激的人类舌下腺和颌下腺唾液中的复杂蛋白质混合物通过二维（2-D）凝胶电泳进行分离，经银染可视化，然后通过免疫染色进行分析。通过与特异性抗体和过氧化物酶偶联的二抗孵育（蛋白质印迹法）来鉴定特定蛋白质。电泳和银染显示，在2微升未浓缩的混合物中有50多种蛋白质成分。当使用抗全血清抗体时，蛋白质印迹技术能够检测到血浆来源的蛋白质斑点，但只能发现白蛋白斑点。用特异性抗体鉴定出了白蛋白、分泌型IgA、酸性磷酸酶和α淀粉酶。