Decker R S, Decker M L, Poole A R
J Histochem Cytochem. 1980 Mar;28(3):231-7. doi: 10.1177/28.3.6986433.
Lysosomal cathepsin D has been localized with the electron microscope employing an indirect immunohistochemical method using peroxidase labeled, monospecific antibody Fab' subunits. The acid proteinase has been demonstrated within secondary lysosomes of cardiac myocytes and interstitial cells, but not in components of the Golgi apparatus or endoplasmic reticulum. Incubations with a variety of peroxidatic inhibitors suggests that the staining that is observed in secondary lysosomes is attributable to the peroxidase-labeled antibody and not to endogenous oxidation of DAB. The protocol outlined here provides a reproducible method to localize the major lysosomal acid proteinase of the heart at the subcellular level.
利用过氧化物酶标记的单特异性抗体Fab'亚基,通过间接免疫组织化学方法,在电子显微镜下对溶酶体组织蛋白酶D进行了定位。已在心肌细胞和间质细胞的次级溶酶体内证实了这种酸性蛋白酶的存在,但在高尔基体或内质网的成分中未发现。用多种过氧化物酶抑制剂进行孵育表明,在次级溶酶体中观察到的染色归因于过氧化物酶标记的抗体,而不是DAB的内源性氧化。本文概述的方案提供了一种可重复的方法,用于在亚细胞水平定位心脏主要的溶酶体酸性蛋白酶。