Morenkov O S, Mantsygin Iu A
Tsitologiia. 1990;32(12):1225-30.
Monoclonal antibodies (MAb) to the thymidine analog 5-bromo-2'-deoxyuridine (BrdUrd) and to horseradish peroxidase (HRPO) have been produced and characterized. On the basis of Mab to HRPO, complexes of the antibodies and HRPO (PAP-complexes) for immunochemical investigations are prepared. The possibility to identify proliferating cells in cultures of mouse myeloma Sp2/0 and mouse fibroblasts NIH 3T3 using Mab to BrdURd by the PAP-method is shown. The conditions of performing the analysis were optimized. The effect of various techniques ot cell fixation and cell DNA denaturation on cell morphology and on specific staining of nuclei of BrdUrd-containing cells in investigated.
已制备并鉴定了针对胸苷类似物5-溴-2'-脱氧尿苷(BrdUrd)和辣根过氧化物酶(HRPO)的单克隆抗体(MAb)。基于针对HRPO的单克隆抗体,制备了用于免疫化学研究的抗体与HRPO的复合物(PAP复合物)。展示了通过PAP法使用针对BrdURd的单克隆抗体在小鼠骨髓瘤Sp2/0和小鼠成纤维细胞NIH 3T3培养物中鉴定增殖细胞的可能性。对进行分析的条件进行了优化。研究了各种细胞固定和细胞DNA变性技术对细胞形态以及对含BrdUrd细胞细胞核特异性染色的影响。