HANNIBAL M J, NACHLAS M M
J Biophys Biochem Cytol. 1959 Mar 25;5(2):279-88. doi: 10.1083/jcb.5.2.279.
This report describes additional studies of the lyo and desmo components of esterase, alkaline phosphatase, acid phosphatase, leucine aminopeptidase, and beta-glucuronidase. The techniques used have already been reported (7). Enzyme diffusion occurs to different degrees in different fixatives, and varies somewhat with each enzyme. Loss of enzymatic activity during fixation occurs as a result of both inactivation due to the chemical reaction of the fixative with the enzymic protein, and diffusion of the lyo component into the fixative. The amount of diffusion into formalin can be reduced by the addition of salts, sucrose, or methocel. The pH of the aqueous medium significantly influences the removal of the lyo fraction from the tissue section. A striking similarity can be noted in the proportions of each fraction of enzyme present in the kidney of the rat, dog, and man. The procedure of fixation and paraffin embedding of tissue blocks does not wholly prevent the diffusion of the lyo component from the tissue sections when they are subsequently immersed in the aqueous incubation medium.
本报告描述了对酯酶、碱性磷酸酶、酸性磷酸酶、亮氨酸氨肽酶和β-葡萄糖醛酸酶的冻干和桥粒成分的进一步研究。所使用的技术已在之前报道过(7)。酶扩散在不同固定剂中程度不同，且每种酶也略有差异。固定过程中酶活性的丧失是由于固定剂与酶蛋白的化学反应导致的失活以及冻干成分扩散到固定剂中。通过添加盐、蔗糖或甲基纤维素可以减少扩散到福尔马林中的量。水性介质的pH值显著影响组织切片中冻干部分的去除。在大鼠、狗和人的肾脏中存在的每种酶组分的比例可以观察到显著的相似性。当组织块随后浸入水性孵育介质中时，组织块的固定和石蜡包埋过程并不能完全防止冻干成分从组织切片中扩散。
