钙调蛋白磷酸酶（Calcineurin，CN）是目前发现唯一活性受Ca2+/钙调素（Calmodulin，CaM）调节的Ser/Thr蛋白磷酸酶。它由催化亚基A和调节亚基B按1:1的比例紧密结合组成的异二聚体。CN的分子量为80KDa，其中钙调蛋白磷酸酶A亚基（CNA）的分子量为61KDa，钙调蛋白磷酸酶（CNB）的分子量为19KDa。CN在免疫调节和信号转导等方面都有重要作用，而且它还与阿尔茨海默病（Alzheimer disease，AD）、心肌肥大以及自身免疫病如类风湿性关节炎等疾病的发生有关。CNB是CN的调节亚基，它的分子量小，性质稳定，CNB除作为CNA的调节亚基外，本实验室研究发现CNB还具调节天然免疫和抗肿瘤作用。近年来发现，在外界刺激或体液因子（如AngⅡ、ET-1）导致细胞内Ca2+ 浓度升高，激活CN，使NFAT3去磷酸化入核，诱导心肌肥大。缺血再灌的大鼠模型基因芯片分析结果显示，心脏梗塞区CNB表达量较非梗塞区下降明显，这一结果更直接提示了CNB在心脏缺血再灌中必然发挥某种作用。CN分子包括CNB在心肌的表达量非常丰富。这些结果都提示CNB有可能在心脏缺血再灌损伤中发挥一定的作用。本实验是通过进行大鼠心肌细胞的原代和传达培养，建立大鼠心肌细胞缺氧/复氧损伤模型，探寻CNB对大鼠心肌细胞缺氧/复氧损伤凋亡的影响。本实验室在制备低纯度CNA多克隆抗体时遇到困难，通过实验，发明了一种用于制备多克隆抗体的新方法，即用含有一种共同目的抗原的两种或两种以上的抗原去对同一动物进行两次或两次以上的免疫，经过免疫的动物对共同的目的抗原的多次免疫应答，产生仅识别共同目的抗原的高特异性抗体IgG。这个方法不仅适用于实验室制备蛋白质抗原的多克隆抗体，也可应用于其他类型的抗原，甚至可以用于单克隆抗体的制备过程。

Calcineurin (CN) is a unique Ca2+/calmodulin (CaM)-stimulated Ser/Thr protein phosphatase. It is a heterodimeric enzyme consisting of a catalytic A subunit (CNA) and regulatory B subunit (CNB). Its MW is about 80 KDa, with a 61KDa CNA and a 19KDa CNB. Calcineurin (CN) plays an important role in the regulation of immunity and signaling transduction pathway. Besides, it also involves in the genesis of Alzheimer’s Disease, cardiac hypertrophy and auto-immune diseases, e.g. arthritis deformans, and so on. CNB is a small molecule with stable characteristics and can promote the activity of CN. Besides being the CNA regulating subunit, CNB is also found playing role in regulating native immune and anti-tumor function. Recent research demonstrated that the increasing incellular Ca2+ concentrations, caused by the environment stimulates and humoral factors (such as Ang II, ET-1), lead to the activation of CN, which will dephosphorylate the NFAT3. Then dephosphorylated NFAT3 transferring into cellular nucleus, induce myocardium hypertrophy. Gene chip results of ischemia/reperfusion mouse reveals that the expression level of CNB decrease obviously in myocardial infarction area comparing to the non-infarction area. In addition, CN including CNB are abundant in cardio muscle. These evidences indicate that CNB may function in heart ischemia/reperfusion injury. So we did the experiments of serial subcultivation and primary cultivation for cardiomyocyte, and established the model of after hypoxia/reoxygenation injury for cardiomyocyte. The apoptosis was also quantificationally examined by the newbuilt model.It’s hard to get high valence and high monospecific Polyclonal antibodies(PcAb) against CNA because of its low purity. Our lab has overcome the general obstacle of producing PcAb by a new approach after many experiments. The approach has been optimized for producing PcAb with impure proteins. The approach is based on the principle of the immunoglobulin (Ig) class switch in the immune response. We have used different impure proteins to immunize mice in turn and obtained monospecific PcAb against a single protein. Remarkably, The approach not only offered a simple and general alternative to other methods for producing described previously, but also disclosed a novel process of immunization that could be used to produce monoclonal antibodies (mAb).