摘要
作为一种新型的速效局部止血药和工具酶,凝血酶在临床和生物学研究中的应用十分广泛,牛血浆是其重要的来源之一。等电点沉淀是提取牛凝血酶首要和关键的步骤,测定其等电点后,再用此法时将得到更纯的凝血酶粗制品。本实验的目的是采用载体两性电解质pH梯度等电聚焦电泳的方法,结合SDS-PAGE测定牛凝血酶的等电点。经双向电泳后,SDS聚丙烯酰胺凝胶中出现了4个清晰的斑点,分别测定它们的分子量和等电点,其中一个斑点与牛凝血酶B链的分子量一致为32kDa,其等电点为5．19。
As a new quick-acting partial hemostat and implemental enzyme, thrombin has been widely used in clinical and biological researches. Bovine plasma is one of the important thrombiu resources. PI （=lsoelectric Point） sedimentation is the first and pivotal procedure when abstracting bovine thrombin from plasma. After assaying the isoelectric point of bovine thrombin, the pure semifinished product of thrombin will be obtained. The purpose of the experiment is to assay the isoelectric point of bovine thrombin by carrier ampholytes pH gradient isoelectrofocusing （IEF） electrophoresis and SDS-PAGE. The result of 2D electrophoresis is that four spots, whose molecular weight and isoelectric point can be assayed, from the SDS-polyacrylamide gel clearly, among which, a spot is equivalent to chain B of bovine thrombin in molecular weight （32kDa）, and its isoelectric point is 5.19.
出处
《现代生物医学进展》
CAS
2006年第11期85-88,共4页
Progress in Modern Biomedicine
基金
国家"985工程"专项资助(CUN985-3-3)
