BrdU抗血清制备和应用的研究——Ⅰ.高度特异的BrdU抗血清的制备与特性

半抗原BrU通过与BSA偶联制备了完全抗原,经过光吸收、SDS聚丙烯酰胺凝胶电泳和琼脂糖凝胶电泳的测定表明,核苷-蛋白质复合物符合制备的要求,每个BSA上估计大约平均有10.3个BrU。用常规免疫的方法获得兔抗BrdU的抗血清,与BrU-EA的双向扩散效价高达32。抗血清稀释128万倍时仍可见明显的ELISA阳性反应。与以前所报道的BrdU抗血清不同,该抗血清具有高水平的识别能力,已达到BrdU单克隆抗体的识别水平,无须纯化即可用于染色体及核酸的研究。

Preparation and Utilization of BrdU Antiserum I.Preparation and Characterization of Highly Specific BrdU Antiserum

The hapten BrU was coupled with BSA to produce the complete antigen of BrdU. The complex of BrU and BSA was determined with methods of light absorbance, SDS polyacry-lamide gel electrophoresis and agrose gel electrophoresis. The results showed that the nucleotide-protein complex was accorded with requirements of preparation, averagely about 10.3 BrU per BSA.. The BrdU antiserum of rabbit was reacted with the BrU-EA in double immunodiffu-sion tests, and a stricking percipitation line was found. Its titre was 32. EL1SA reaction was still quite clear when the antiserum was diluted to 1:12800 in PBS. Some characters of this antiserum was differ from other antiserums which had been reported before. This BrdU antiserum was highly specific, similar to the specification of BrdU monoclone antibody. Therefore this anti-serum was not necessary to purify for study of BrdU antibody techniques in the research of chromosomes and DNA.