Danø K, Dabelsteen E, Nielsen L S, Kaltoft K, Wilson E L, Zeuthen J
J Histochem Cytochem. 1982 Nov;30(11):1165-70. doi: 10.1177/30.11.6183313.
A monoclonal antibody against a 52,000 dalton human plasminogen activating enzyme (HPA52) was used for immunofluorescence staining of cultured glioblastoma cells. The fluorescence was located in the cytoplasm of the cells. A pronounced variation in the staining intensity was observed between the individual cells. The specificity of the fluorescent stain was supported by the findings that 1) no staining was obtained with a monoclonal antibody of the same subclass, but with irrelevant specificity (anti-2,4,6-trinitrophenyl); 2) adsorption with HPA52 purified to homogeneity removed the ability of anti-HPA52 to mediate staining; 3) the glioblastoma cells contained HPA52, as measured by enzymatic assay, while melanoma cells that were not stained did not contain HPA52 activity; 4) dexamethasone reduced both the enzymatically determined HPA52 content and the immunofluorescence in parallel, while progesterone affected none of these parameters; 5) we have previously found that culture fluid conditioned by the glioblastoma cells apart from HPA52 does not contain detectable amounts of any protein that binds to anti-HPA52. Several advantages of immunohistochemical detection of plasminogen activators compared with enzyme histochemical methods are discussed, among these that the immunohistochemical method distinguishes between plasminogen activators of different types.
一种针对52,000道尔顿人纤溶酶原激活酶（HPA52）的单克隆抗体用于培养的胶质母细胞瘤细胞的免疫荧光染色。荧光位于细胞的细胞质中。在各个细胞之间观察到染色强度有明显差异。荧光染色的特异性得到以下发现的支持：1）用相同亚类但具有无关特异性的单克隆抗体（抗2,4,6-三硝基苯基）未获得染色；2）用纯化至同质的HPA52吸附消除了抗HPA52介导染色的能力；3）通过酶测定法测量，胶质母细胞瘤细胞含有HPA52，而未染色的黑色素瘤细胞不含有HPA52活性；4）地塞米松同时降低了酶测定的HPA52含量和免疫荧光，而孕酮对这些参数均无影响；5）我们之前发现，除HPA52外，由胶质母细胞瘤细胞条件培养液中未检测到任何与抗HPA52结合的蛋白质。讨论了与酶组织化学方法相比，纤溶酶原激活剂免疫组织化学检测的几个优点，其中包括免疫组织化学方法可区分不同类型的纤溶酶原激活剂。
