钙调素抗体的制备和鉴定

将从猪脑中所得SDS—PAGE纯的钙调素进行化学修饰,得(DNPm—CaM)n—BSA。将此修饰物免疫California白兔得抗血清。钙调素的ELISA测定表明:抗血清与天然牛脑和天然猪脑钙调素、CaBP_(72)(钙结合蛋白,分子量72KDa)、CaN(神经钙蛋白)、TnI(肌钙蛋白抑制亚单位)的交叉反应率分别为100％、100％、0.7％、0.4％、0.02％;与CaBP_(11)、CaBP_(12)、TnT、TnC、S—100、MLC(肌球蛋白轻链)均无交叉反应。免疫转移鉴定技术示:大鼠脑匀浆上清液显示强弱两条区带,强染色带相当于钙调素的位置,弱显色带分子量小于钙调素;其它所测大鼠组织匀浆上清液只在钙调素位置显示单一区带。

PREPARATION AND IDENTIFICATION OF ANTI-CAL MODULIN ANTIBODY

(DNPm-CaM) n-BSA was prepared by modifying the porcine brain ealmodulin(CaM)which had been shown to be homogeneous by SDS-PAGE, and then was used to immune California white rabbits. The antiserum obtained was tested for its immunoreaction by means of ELISA against native bovine brain CaM, native porcine brain CaM, CaBP72(Ca2 binding protein 72KDa ) , CaN ( calcineurin ) and TnI ( inhibitor troponin ) , showing the cross-reaction ratios were 100, 100, 0.7, 0.4 and 0.02%respectively; while no cross-reaction occurred against CaBP11, CaBP12, TnT ( tropotroponin), TnC ( Ca2 combining troponin ) , S100, and MIC ( myosin light chain ) . Immonoblotting technique for CaM demonstrated 2 bands appearing in the panel of the supernatant of homogenized rat brain; one was apparent and occupying a position corresponding to that of standard CaM and the other one was rather faint and representing a molecular weight less than that of CaM. The supernatanes from other rat organs ( liver, testis and heart ) gave just one and the same band as the standard CaM.