Moore B M, Kang B, Flurkey W H
Department of Chemistry, Indiana State University, Terre Haute 47809.
Histochemistry. 1989;90(5):379-81. doi: 10.1007/BF00508316.
Mushrooms were cut into vertical and horizontal sections. These sections were blotted onto nitrocellulose sheets and the sheets were then stained for tyrosinase using L-dopa. Tyrosinase was localized throughout the mushroom tissues but more enzyme was located in the epidermis of the cap, the gill region, and the stipe. Preincubation of the nitrocellulose sheets in specific inhibitors of tyrosinase completely blocked enzyme staining, suggesting that the enzyme stained areas on the nitrocellulose blots were regions of tyrosinase activity. Immunochemical localization of tyrosinase was similar to that observed by histochemical staining. Nitrocellulose blotting of mushrooms allows localizations of enzyme at the whole tissue level and may be useful for other enzymes in mushrooms as well.
将蘑菇切成垂直和水平切片。将这些切片印迹到硝酸纤维素膜上，然后使用L-多巴对膜进行酪氨酸酶染色。酪氨酸酶定位于整个蘑菇组织中，但更多的酶位于菌盖表皮、菌褶区域和菌柄中。在酪氨酸酶的特异性抑制剂中预孵育硝酸纤维素膜可完全阻断酶染色，这表明硝酸纤维素印迹上的酶染色区域是酪氨酸酶活性区域。酪氨酸酶的免疫化学定位与组织化学染色观察到的结果相似。蘑菇的硝酸纤维素印迹法可在整个组织水平上定位酶，也可能对蘑菇中的其他酶有用。
