SDS—PAGE分离不孕奶牛宫颈黏液AsAb

摘要

免疫不孕严重影响奶牛正常繁殖，相对于其他不孕类型不容易被确诊，准确对孕牛与不孕牛之间宫颈黏液AsAb的差异进行定性和定量分析，对奶牛免疫不孕的治疗研究有重要的意义。本试验将浅盘凝集检测和实际生产结果相结合确定的不孕奶牛和可孕奶牛作为研究对象，对宫颈黏液中蛋白进行SDS—PAGE电泳并对其分离条件进行优选。结果表明，本试验建立了一种在8％分离胶上，以15p~L上样量，100V，电泳3～4h的SDS—PAGE分离奶牛宫颈黏液的有效方法：对浅盘凝集检测和实际生产记录相结合确定AsAb免疫性不孕奶牛的宫颈黏液，用%Immune infertility can affect the normal reproduction of dairy cows, seriously and it is more difficult to diagnosis it than other types of infertility. It is very important that the quantitative and qualitative analysis for the difference of AsAb between fertility and infertility cows were researched. Proteins in cervical mucus of the infertility cows and fertility cows were determined by TAT test combining with actual production results, and separated by SDS-PAGE. All of conditions were optimized. Result showed that a kind of effective method of separation of protein in dairy eows＇s cervical mucus by SDS-PAGE was established. The optimizing conditions were 8% separation glue, 15p~L sample, 100V, electrnphoresis for 3-4h. A protein, with the molecular weight of 22-25kDa, which preliminarily determined is AsAb Light Chain, was gotten. Infertility cows contained AsAb Light Chain in cervical mucus with 100% （24/24）, and the percentage of fertility cows containing AsAb Light Chain in cervical mucus was 14.3% （2/14）, but the expression level of AsAb Light Chain was lower than that of infertility cows.