The whole aspect elucidation of gap junction formation process between adjecent cells using the anti-connexin 32 monoclonality antibody.

1.识别Cx32胞外表位的HAM12抗体的制备合成的多肽与弗氏完全佐剂混合免疫BALB/c小鼠。该多肽与p160-172的连接蛋白32胞外第二环相同，半胱氨酸位置用白喉类毒素标记。小鼠体内浆细胞产生抗体后，NS-1骨髓瘤细胞与免疫小鼠的脾细胞进行细胞融合，杂交瘤细胞正在克隆中。分析大鼠肝细胞Cx32在体内的形成过程。将注射HAM8抗体的大鼠肝脏冰冻切片仅用FITC标记的抗鼠IgG染色，通过荧光显微镜观察切片，根据形成速度估计缝隙连接斑点的形成过程，并提出假设。详情见Anat Rec262：147152,20013。体外培养大鼠肝细胞Cx32形成过程的分析。通过分离培养肝细胞的原代培养方法完成，该成果是在目前倒置显微扩增技术的差示干涉荧光显微镜下进行的。

1. The production of HAM12 antibody hich recognizes Cx32 extacellular epitope. BALB/c mice were immnunaized by the mixture of syuthetized peptides and Freunt Complete Adjuvant. The peptide was identical to the 2nd extracellular loop of connexin 32 from p160-172 and it was labeled with diphtheria toxoid at the Cys position.After chicking whethere the plasma cell in the mouse were producing the antibody, cell fusion of NS- 1 myeloma cells and spleen cells from immunized mouse was done, then the hybridomas are under cloning now.2. The analysis of the Cx32 formation process of the rat hepatocyte in vivo.The liver frozen sections from HAM8 antibody injected rats were staind with only FITC-labeled anti-mouse IgG.As a result of observing the slice by fluorescence microscope, the formation process of the gap junction spot was estimated at the formation speed, and the hypothesis was made. The details were appeared Anat Rec 262 : 147-152, 20013. The analysis of the Cx32 formation process of the rat hepatocyte in vitro.Though primary culture method of the dissociated heachievement-finished, the achievement is being carried out by conformer and under the differential interference fluorescence microscope which inverts the micromanuplation technique at present too.