Karsten U, Butschak G, Cao Y, Goletz S, Hanisch F G
Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany.
Hybridoma. 1995 Feb;14(1):37-44. doi: 10.1089/hyb.1995.14.37.
A new monoclonal antibody to the Thomsen-Friedenreich (TF) antigen (or, more precisely, epitope; Gal beta 1-3GalNAc-) has been developed that is specific for both anomeric forms of this disaccharide (TF alpha and TF beta, including related structures on glycolipids), and not assay restricted. We demonstrate that this avid antibody (A78-G/A7) is well suited for immunohistochemistry on paraffin-embedded and cryosectioned tissues, immunoblotting, ELISA techniques, and hemagglutination. Immunohistochemistry on paraffin sections does not require proteolytic or microwave pretreatment. The binding characteristics of this antibody are largely independent of variations in pH (6.0-8.2) and temperature (4-37 degrees C). Immunoblotting with KG-1 (human acute myelogenous leukemia) cells revealed a series of TF-active glycoproteins with a main band at about 155 kDa. Immunoprecipitation was performed using a new technique applicable to IgM-type antibodies.
一种针对汤姆森-弗里登赖希（TF）抗原（或者更准确地说是表位；Galβ1-3GalNAc-）的新型单克隆抗体已被研发出来，它对这种二糖的两种异头物形式（TFα和TFβ，包括糖脂上的相关结构）具有特异性，且不受检测限制。我们证明这种高亲和力抗体（A78-G/A7）非常适合用于石蜡包埋组织和冷冻切片组织的免疫组织化学、免疫印迹、酶联免疫吸附测定（ELISA）技术以及血细胞凝集实验。石蜡切片的免疫组织化学不需要进行蛋白水解或微波预处理。该抗体的结合特性在很大程度上不受pH值（6.0 - 8.2）和温度（4 - 37摄氏度）变化的影响。用KG - 1（人急性髓性白血病）细胞进行免疫印迹显示出一系列TF活性糖蛋白，其中主要条带位于约155 kDa处。免疫沉淀采用了一种适用于IgM型抗体的新技术进行。
