Narisawa S, Harmey D, Magnusson P, Millán J L
The Burnham Institute, La Jolla, CA 92037, USA.
Tumour Biol. 2005 May-Jun;26(3):113-20. doi: 10.1159/000086482. Epub 2005 Jun 17.
A panel of 19 monoclonal antibodies (MAbs) against human tissue-nonspecific (liver/bone/kidney) alkaline phosphatase (TNAP) was obtained through the ISOBM TD-9 workshop. In the present study, the reactivity of these MAbs has been characterized against mouse TNAP. A mouse embryonic stem cell line, frozen sections of long bones, alkaline phosphatase extracted from mouse bone, and serum were used as the source of TNAP for individual assays. Each MAb was tested for immunoreactivity to mouse TNAP by Western blot analysis, immunohistochemistry and enzyme immunoassay. Antibodies 314 and 315 reacted strongly with mouse TNAP in Western blots, while all other antibodies were negative. By immunohistochemistry, antibodies 314, 315 and 333 produced strong positive staining using frozen sections, while antibody 334 was moderately positive. Enzyme immunoassays indicated that MAb 333 was also able to bind to serum TNAP. These antibodies represent very useful reagents to study the pathophysiological expression of TNAP in mouse tissues and in mouse serum.
通过国际单克隆抗体标准品库（ISOBM）TD-9研讨会获得了一组针对人组织非特异性（肝脏/骨骼/肾脏）碱性磷酸酶（TNAP）的19种单克隆抗体（MAb）。在本研究中，已对这些单克隆抗体针对小鼠TNAP的反应性进行了表征。将小鼠胚胎干细胞系、长骨冰冻切片、从小鼠骨骼中提取的碱性磷酸酶以及血清用作各个检测中TNAP的来源。通过蛋白质免疫印迹分析、免疫组织化学和酶免疫测定法检测每种单克隆抗体对小鼠TNAP的免疫反应性。抗体314和315在蛋白质免疫印迹中与小鼠TNAP强烈反应，而所有其他抗体均为阴性。通过免疫组织化学，抗体314、315和333使用冰冻切片产生强阳性染色，而抗体334呈中度阳性。酶免疫测定表明单克隆抗体333也能够结合血清TNAP。这些抗体是研究TNAP在小鼠组织和小鼠血清中的病理生理表达的非常有用的试剂。
