Mason D Y, Sammons R
J Immunol Methods. 1978;20:317-24. doi: 10.1016/0022-1759(78)90267-3.
Soluble immune complexes of horseradish peroxidase and antibody to peroxidase (PAP) have been widely used in the 'unlabelled antibody' method for the immunocytochemical detection of cellular antigens. This paper describes a simple and rapid method for preparation of these complexes by column chromatography of a mixture of the enzyme and the IgG fraction of antiperoxidase antiserum on Sephacryl S-200. The material eluting in the void column consists of stable soluble PAP complexes, with a molar peroxidase: antiperoxidase ratio of 0.8 and a molecular weight of approximately 400,000. When tested immunocytochemically this material gives identical results to those obtained with conventionally prepared PAP.
辣根过氧化物酶与抗过氧化物酶抗体（PAP）的可溶性免疫复合物已广泛应用于“未标记抗体”法，用于细胞抗原的免疫细胞化学检测。本文描述了一种简单快速的方法，通过在Sephacryl S - 200上对酶和抗过氧化物酶抗血清的IgG组分混合物进行柱色谱来制备这些复合物。在空柱中洗脱的物质由稳定的可溶性PAP复合物组成，过氧化物酶与抗过氧化物酶的摩尔比为0.8，分子量约为400,000。当进行免疫细胞化学测试时，该物质给出的结果与用传统方法制备的PAP所获得的结果相同。
