Theoharides T C, Canellakis Z N
J Biol Chem. 1976 Mar 25;251(6):1781-4.
Specific antiserum to a purified preparation of ornithine decarboxylase was prepared and successfully labeled with 125I. Monospecificity of this antiserum was attained by repeated precipitation with normal rat liver. The conditions for optimal antigen/antibody reaction were investigated. When the antibody was tested against either crude or purified regenerating rat liver enzyme a single precipitin line was observed both with Ouchterlony double diffusion plates and with immunoelectrophoresis. The specificity of the purified antiserum was also evaluated by sodium dodecyl sulfate gel electrophoresis of the 14C-labeled enzyme.antibody complex, the radioactivity of which appeared as a sharp peak in the 90,000 molecular weight region. Finally, an approximate antigen/antibody molar ratio was determined using labeled enzyme and labeled antiserum.
制备了针对纯化的鸟氨酸脱羧酶制剂的特异性抗血清,并用125I成功标记。通过用正常大鼠肝脏反复沉淀获得了该抗血清的单特异性。研究了最佳抗原/抗体反应条件。当用粗制或纯化的再生大鼠肝脏酶检测抗体时,在双向琼脂扩散平板和免疫电泳中均观察到一条单一的沉淀线。还通过对14C标记的酶-抗体复合物进行十二烷基硫酸钠凝胶电泳来评估纯化抗血清的特异性,其放射性在90,000分子量区域呈现为一个尖锐峰。最后,使用标记的酶和标记的抗血清确定了近似的抗原/抗体摩尔比。