Nunn D J, Mezei C
J Neurochem. 1984 Jan;42(1):158-65. doi: 10.1111/j.1471-4159.1984.tb09712.x.
To explore the immunological properties of PO protein, antibodies were elicited in rabbits against the purified chick PO protein. Peripheral nervous system protein was fractionated on sodium dodecyl sulfate-polyacrylamide slab gels and then transferred electrophoretically ("blotted") onto nitrocellulose sheets. The PO protein was detected by its capacity to bind its specific antibody present in the rabbit serum. The PO-specific antibody complex was then exposed to goat anti-rabbit immunoglobulin G (IgG) coupled to peroxidase or labeled with 125I. The resulting PO antigen-antibody "sandwich" was visualized and quantitated by densitometry of the colored peroxidase reaction product or by autoradiography and gamma-radiation counting of the 125I-IgG complex. The methods permitted quantitation of the PO protein in various nerve extracts. The limit of detection of the PO antigen was about 1 ng of protein. The antibody was specific for the PO glycoprotein in the peripheral nerve extracts. The PO proteins from various species, including human, were also detected by the antibody to chick PO protein. Preliminary experiments indicate the solid-phase immunoassay is a useful method for monitoring PO protein levels in small quantities of tissue extracts under various physiological and pathological conditions.
为了探究PO蛋白的免疫学特性，用纯化的鸡PO蛋白在兔体内诱导产生抗体。将外周神经系统蛋白在十二烷基硫酸钠-聚丙烯酰胺平板凝胶上进行分级分离，然后通过电泳转移（“印迹”）到硝酸纤维素膜上。通过PO蛋白与兔血清中存在的特异性抗体结合的能力来检测PO蛋白。然后将PO特异性抗体复合物与偶联过氧化物酶或用125I标记的山羊抗兔免疫球蛋白G（IgG）接触。通过对有色过氧化物酶反应产物进行光密度测定，或通过对125I-IgG复合物进行放射自显影和γ射线计数，对产生的PO抗原-抗体“夹心”进行可视化和定量分析。这些方法能够对各种神经提取物中的PO蛋白进行定量。PO抗原的检测限约为1 ng蛋白。该抗体对外周神经提取物中的PO糖蛋白具有特异性。针对鸡PO蛋白的抗体也能检测到包括人类在内的各种物种的PO蛋白。初步实验表明，固相免疫测定是一种在各种生理和病理条件下监测少量组织提取物中PO蛋白水平的有用方法。
