Aumüller G, Vedder H, Enderle-Schmitt U, Seitz J
Department of Anatomy and Cell Biology, Philipps-Universität, Marburg, Federal Republic of Germany.
Prostate. 1987;11(1):1-15. doi: 10.1002/pros.2990110102.
Acid phosphatase (E.C. 3.1.3.2.) has been isolated from canine prostatic gland homogenates by gel permeation chromatography (AcA34 or G150), by affinity chromatography (con A-Sepharose), or by using fluid phase liquid chromatography (FPLC) using Superose 12 and Mono P columns. Acid phosphatase-enriched fractions were submitted to analytical SDS-PAGE or to analytical isoelectric focusing. A protein with a molecular weight of 30 kD (on SDS gels) was used for immunization of rabbits. The antiserum produced was cross-reactive with prostatic acid phosphatase (canine and human) as shown by immunoblotting. When applied to paraffin or plastic sections of normal canine prostate, a positive immunoreaction was found exclusively in the secretory cells. In experimentally altered glands (castration and/or hormone treatment), a varying pattern of immunoreactive cells was found. In canine prostatic carcinomas, intensively reacting cell clusters were found along with nonreactive cells. The antiserum was also slightly cross-reactive with the respective human antigen, but the cross-reactivity of an antiserum prepared against human prostatic secretory acid phosphatase with canine prostatic acid phosphatase was far more pronounced.
酸性磷酸酶(E.C. 3.1.3.2.)已通过凝胶渗透色谱法(AcA34或G150)、亲和色谱法(伴刀豆球蛋白A-琼脂糖凝胶)或使用Superose 12和Mono P柱的液相色谱法(FPLC)从犬前列腺匀浆中分离出来。富含酸性磷酸酶的组分进行了SDS-PAGE分析或等电聚焦分析。一种分子量为30 kD(在SDS凝胶上)的蛋白质用于免疫兔子。免疫印迹显示,产生的抗血清与前列腺酸性磷酸酶(犬和人)有交叉反应。当应用于正常犬前列腺的石蜡或塑料切片时,仅在分泌细胞中发现阳性免疫反应。在实验性改变的腺体(去势和/或激素治疗)中,发现了不同模式的免疫反应性细胞。在犬前列腺癌中,发现了强烈反应的细胞簇以及无反应的细胞。该抗血清与相应的人类抗原也有轻微交叉反应,但针对人类前列腺分泌性酸性磷酸酶制备的抗血清与犬前列腺酸性磷酸酶的交叉反应更为明显。