等电聚焦电泳方法的建立和牛凝血酶等电点的初步测定

s a new quick result partial astringent and implemental enzyme, thrombin is used in clinical and biological research quite widely. Bovine plasma is one of the important thrombin resources. pI sedimentation is the first and pivotal procedure when abstract bovine thrombin form plasma. After identify the isoelectric point of bovine thrombin, the preliminary manufacture of thrombin will be more rarefied. The purpose of the experiment is to establish the methodology and procedure of carrier amphlytes pH gradient isoelectrofocusing (IEF) electrophoresis according identify the isoelectric point (pI) of trypsin and apply the method biding SDS-PAGE to identify the isoelectric point of bovine thrombin. The result of 2D electrophoresis is that four spots whose molecular weight and isoelectric point can be identified emerged in the SDS-polyacrylamide gel clearly, and among them, a spot equivalent to chain B of bovine thrombin in molecular weight (32kDa) is the one we wanted. Therefore, the isoelectric point of bovine thrombin is 5.19.