Isoelectric focusing and immunoblotting analysis of thyroglobulin from different thyroid diseases.

Thyroidology Pub Date : 1991-01-01
A Olivieri, A Marzullo, G B Salabé, E Gilardi, C Fazzini, S Carta
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Abstract

Thyroglobulins (Tgs) have been obtained from macro-microfollicular goiter, toxic adenomas, papillary carcinomas and metastatic lympho-node and chromatographed by gel filtration. The 19S Tgs so obtained have been characterized by isoelectric focusing (IEF) and immunoblotting. IEF showed microheterogeneity pattern of Tgs from normal tissue, macro-microfollicular goiter and toxic adenomas to be within pH range from 4.2 to 4.5, while that from papillary carcinomas and metastatic lympho-node showed a wider microheterogeneity. Passive blotting of focused Tgs and immunoreaction with rabbit anti-Tg peroxidase conjugated antibody have been carried out and a positive reaction for all examined samples have been evidenced. Also two bands, focused at more cathodic pH value (pH 4.7), observed in IEF patterns of Tgs from carcinomas and metastatic lympho-node, gave a positive reaction with anti-Tg conjugated antibody. These differences observed in IEF and immunoblotting patterns of Tgs from papillary carcinomas and metastatic lympho-node are discussed in this report.

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不同甲状腺疾病甲状腺球蛋白的等电聚焦及免疫印迹分析。
甲状腺球蛋白(Tgs)已从大、微滤泡性甲状腺肿、中毒性腺瘤、乳头状癌和转移性淋巴结中获得,并经凝胶过滤进行了层析。用等电聚焦(IEF)和免疫印迹技术对所获得的19S Tgs进行了表征。IEF显示正常组织、大-微滤泡性甲状腺肿和毒性腺瘤的Tgs在pH值4.2 ~ 4.5范围内具有显微异质性,而乳头状癌和转移性淋巴结的Tgs具有更广泛的显微异质性。用兔抗tg过氧化物酶偶联抗体对聚焦的tg进行被动印迹和免疫反应,所有检测样品均呈阳性反应。此外,在癌和转移性淋巴结的Tgs的IEF模式中观察到两个条带,集中在更阴极的pH值(pH 4.7),抗tg偶联抗体呈阳性反应。在乳头状癌和转移性淋巴结的Tgs的IEF和免疫印迹模式中观察到的这些差异在本报告中进行了讨论。
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