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人天冬氨酸蛋白酶A:表达、免疫化学检测及组织定位

Human napsin A: expression, immunochemical detection, and tissue localization.

作者信息

Schauer-Vukasinovic V, Bur D, Kling D, Grüninger F, Giller T

机构信息

F. Hoffmann-La Roche Ltd., Pharma Division, Preclinical Research, Grenzacherstrasse 124, CH-4070, Basel, Switzerland.

出版信息

FEBS Lett. 1999 Nov 26;462(1-2):135-9. doi: 10.1016/s0014-5793(99)01458-1.

Abstract

A novel aspartic proteinase, called napsin, has recently been found in human and mouse. Due to high similarity with cathepsin D a structural model of human napsin A could be built. Based on this model a potential epitope SFYLNRDPEEPDGGE has been identified, which was used to immunize rabbits. The resulting antibody was employed in monitoring the expression of recombinant human napsin A in HEK293 cell line. Western blot analysis confirmed the specificity of the antibody and showed that human napsin A is expressed as a single chain protein with the molecular weight of approximately 38 kDa. Immunohistochemical studies revealed high expression levels of napsin A in human kidney and lung but low expression in spleen.

摘要

最近在人和小鼠中发现了一种名为 napsin 的新型天冬氨酸蛋白酶。由于与组织蛋白酶 D 高度相似,因此可以构建人 napsin A 的结构模型。基于该模型,鉴定出一个潜在的表位 SFYLNRDPEEPDGGE,并用其免疫兔子。所得抗体用于监测重组人 napsin A 在 HEK293 细胞系中的表达。蛋白质印迹分析证实了该抗体的特异性,并表明人 napsin A 以单链蛋白形式表达,分子量约为 38 kDa。免疫组织化学研究显示,napsin A 在人肾和肺中高表达,而在脾中低表达。

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