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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。
The SDS-PAGE pure calmodulin obtained from porcine brain was chemically modified to give (DNPm-CaM) n-BSA. The modified white rabbit was immunized against antisera. The calmodulin ELISA assay showed that antisera reacted with natural bovine and porcine calmodulin, CaBP 72 (calbindin, molecular weight 72 KDa), CaN (calcineurin), TnI (troponin I subunit ) Were 100%, 100%, 0.7%, 0.4% and 0.02%, respectively. The rates of cross-reactivity with CaBP 11, CaBP 12, TnT, TnC, S-100 and MLC No cross-reaction. Identification of immunostaining technology showed: the rat brain homogenate supernatant showed two bands of strong and strong bands corresponding to the position of calmodulin, weak band with molecular weight less than calmodulin; other measured rat tissue homogenization The supernatant shows only a single zone at the calmodulin site.