摘要
本试验对绵羊垂体前叶催乳素进行了提纯和生物学、生物化学及免疫学方面的鉴定.516g 羊全垂体经硫酸铵、乙醇等的逐级提取,最后经 DEAE-纤维素柱层析得到催乳素纯品450mg.用鸽嗉囊局部皮内微量注射法测得其生物学活性为27国际单位(27IU/mg)。PAG 电泳表明有三条带。SDS 电泳测得其分子量为23000。IEFP 电泳表明平均等电点为5.85。用 Dansyl-Cl 法测定其 N-末端氨基酸,只显出苏氨酸一点。表明提纯的催乳素是高纯的。用该催乳素免疫家兔,九周后用琼脂双扩散法测抗血清,其效价最高达1:32,说明该纯化品具有很强的免疫原件.
Through a series of extraction and precipitation with ammonium sulfate and chro- matography on DEAE-cellulose,we achieved 450rag purified prolactin from 516g sheep whole frozen pituitary.The yield was 0.87g per kilogram frozen pituitary. With the method of local micro-introdermal injection over the pigeon crop sac, we deduced that our sample possessed an activity of 27IU/per rag.On the polycry- lamide gel electrophoresis,the sample ran into three distinct bands.The SDS- PAG electrophoresis showed that the molecular weight was about 23,000.The isoelectro focusing electrophoresis showed that the main isoelectric point was 5.85.In the N-terminal amino acid analysis,with the Dansyl--chloride method, only threonine was identified.From all of these,we concluded that our sample washighly purified.Having immunized the rabbits with our sample,we obtainep antisera 9 weeks later,its titre reached 1:32 with agar gel double diffusion method.
关键词
羊
催乳素
制备
提纯
鉴定
抗血清
sheep prolactin
isolation
characterization
antisera
