Isolation and characterization of six monoclonal antibodies raised against human RAD51 recombinant protein
Isolation and characterization of six monoclonal antibodies raised against human RAD51 recombinant protein
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DOI：
10.1089/hyb.1996.15.205
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发表时间：
1996-06-01
期刊：
影响因子：
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通讯作者：
Sturzbecher, HW
中科院分区：
文献类型：
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作者：
Buchhop, S;Donzelmann, B;Sturzbecher, HW
制备了抗重组人RAD 51重组蛋白的单克隆抗体。从无血清细胞培养基中分离IgG亚类的抗体，并通过蛋白A-Sepharose亲和层析纯化。所述抗体可用于在总细胞裂解物的免疫印迹上特异性检测RAD 51蛋白。天然RAD 51蛋白质从人细胞的裂解物中特异性沉淀。此外，这些抗体通过免疫荧光染色容易地检测细胞核中的RAD 51。跨越人RAD 51的完整一级氨基酸序列的重叠肽上的表位作图显示，三个单克隆抗体识别RAD 51上非常接近蛋白质的N-末端（氨基酸16至20）的表位;其他三个单克隆抗体与人RAD 51的氨基酸85至95相互作用。
Monoclonal antibodies were produced against recombinant human RAD51 recombination protein. The antibodies of IgG subclasses were isolated from serum-free cell culture medium and purified by affinity chromatography on protein A-Sepharose. The antibodies can be used to detect specifically RAD51 protein on immunoblots of total cell lysates. Native RAD51 protein is specifically precipitated from lysates of human cells. In addition, these antibodies readily detect RAD51 in the cell nucleus by immunofluorescence staining. Epitope mapping on overlapping peptides spanning the complete primary amino acid sequence of human RAD51 revealed that three monoclonals recognize an epitope on RAD51 very close to the N-terminus of the protein (amino acids 16 to 20); the other three monoclonals interact with amino acids 85 to 95 of human RAD51.