抗rmNDPK-B抗体的制备、纯化、鉴定及应用

为制备抗rmNDPK-B多克隆抗体,以纯化后的rmNDPK-B为抗原,改良法免疫新西兰兔,琼脂双向扩散法分析抗体滴度,SDS-PAGE和Western-blot鉴定纯化后的抗体,结果发现,已获得高效价、高纯度的抗rmNDPK-B多克隆抗体;再分别以此抗体和抗rhNDPK-B抗体为一抗,Western-blot和免疫组织化学法检测小鼠子宫内膜组织中的NDPK-B的表达和分布.结果提示,在检测妊娠小鼠子宫内膜组织NDPK-B的表达时,抗rmNDPK-B抗体的特异性明显高于抗rhNDPK-B抗体.妊娠小鼠子宫内膜组织NDPK-B的表达显著高于非妊娠鼠.这表明用改良法制备的抗rmNDPK-B多克隆抗体具有高特异性、高效价和较高的灵敏性.nm23-M2/NDPK-B可能参与了胚泡着床过程.

Preparation, Purification, Identification of Anti-rmNDPK-B And it's Functional Research

To prepare polyclonal antibody rmNDPK-B, immunized rabbits by the purified protein rmNDPK- B with improved method. Detected the antibody titer by double diffusion test, identified it by SDS-PAGE and Western-blot. The result shows the high-titer and high purified polyclonal antibody rmNDPK-B is obtained. Used antibody rmNDPK-B and antibody rhNDPK-B to tested the distribution of NDPK-B in mice endometrium by immunohistochemical and Western-blot. Compared they differenced. The specificity of antibody rmNDPK-B is higher than antibody rhNDPK-B when the expression Of the NDPK-B in endometria in mice. Furthermore the expressions of pregnant mice are higher than that of non-pregnant mice. This show antibody rmNDPK-B has more specificity, higher titer and more sensitivity. And nm23-M2/NDPK-B may have some function during embryo implantation.