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在福尔马林固定石蜡包埋组织上使用两种不同抗原修复方法对免疫组织化学方法进行优化:63种标志物的经验

Optimization of immunohistochemical methods using two different antigen retrieval methods on formalin-fixed paraffin-embedded tissues: experience with 63 markers.

作者信息

Ramos-Vara J A, Beissenherz M E

机构信息

Veterinary Medical Diagnostic Laboratory, University of Missouri, Columbia 65211, USA.

出版信息

J Vet Diagn Invest. 2000 Jul;12(4):307-11. doi: 10.1177/104063870001200402.

Abstract

Formalin fixation produces cross-links between the proteins and the fixative that alter the ability of some antibodies to recognize antigens. We used formalin-fixed, paraffin-embedded tissues to compare two different antigen retrieval methods for 63 antibodies used in the diagnosis of infectious and neoplastic diseases of animal species. Eighty-four percent of the antibodies needed some type of antigen retrieval for optimal results. Of those antibodies, 67.7% were monoclonal and 32.3% were polyclonal. Steam heat was the method of choice for 31 antibodies. Ten antibodies reacted only with steam heat, but 9 antibodies did not react when steam heat was used. Optimal results were obtained with enzyme digestion for 22 antibodies. Only 10 antibodies yielded optimal results without antigen retrieval; 64% of these antibodies were polyclonal. All antibodies against cytokeratins were optimally retrieved with proteinese K. Antigen retrieval appears to be necessary for the majority of antibodies when used with formalin-fixed, paraffin-embedded tissues.

摘要

福尔马林固定会在蛋白质和固定剂之间产生交联,从而改变某些抗体识别抗原的能力。我们使用福尔马林固定、石蜡包埋的组织,比较了两种不同的抗原修复方法,用于诊断动物物种的感染性和肿瘤性疾病的63种抗体。84%的抗体需要某种类型的抗原修复才能获得最佳结果。在这些抗体中,67.7%是单克隆抗体,32.3%是多克隆抗体。蒸汽加热是31种抗体的首选方法。10种抗体仅与蒸汽加热反应,但9种抗体在使用蒸汽加热时不反应。22种抗体通过酶消化获得了最佳结果。只有10种抗体在没有抗原修复的情况下产生了最佳结果;这些抗体中有64%是多克隆抗体。所有针对细胞角蛋白的抗体都用蛋白酶K进行了最佳修复。当与福尔马林固定、石蜡包埋的组织一起使用时,大多数抗体似乎都需要进行抗原修复。

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