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首页> 外文期刊>Research communications in molecular pathology and pharmacology >Extraction of cytokeratin from the human submandibular gland and its electrophoretic analysis.
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Extraction of cytokeratin from the human submandibular gland and its electrophoretic analysis.

机译：从人下颌下腺中提取细胞角蛋白及其电泳分析。

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摘要

We evaluated a method for extracting cytokeratin (CK) from the normal human submandibular gland and analyzed CK distribution by one- and two-dimensional electrophoresis. Four submandibular glands without histological changes under a light microscope were used as specimens. Intermediate filamentous protein was extracted by a prior modified method CK was not adequately extracted in 1-2% sodium dodecyl sulfate -5% 2-mercaptoethanol solution. On the other hand, 11 bands with molecular weights of 40-58 K were obtained after extraction in 9.5 M urea -5% 2-mercaptoethanol solution. Although fragmentation of cytoskeletal protein was observed, there were no differences in bands according to the strength of homogenization. This fragmentation seemed to be due to restriction degradation by protease. Immunoblotting revealed the reaction of this CK extract to 2 pan-CK antibodies, i.e., K8.13, which recognizes type II basic CK, and C-11, which recognizes CK4, CK5, CK6, CK8, CK10, CK13, CK18. Among monospecific antibodies, anti-CK7 (LDS-68), CK8 (M20), CK13 (KS-1A3), CK18 (CY-90), and CK19 (BA17) antibodies showed positive reactions. Glial fibrillary acidic protein, neurofilament (NF) 68 K, NF160 K, or NF200 K showed no reactions. These results of one- and two-dimensional electrophoretic analysis suggest the presence of CK5, CK7, CK8, CK13, CK14, CK15, CK17, CK18, and CK19 in the normal submandibular gland.
机译：我们评估了一种从正常人下颌下腺中提取细胞角蛋白（CK）的方法，并通过一维和二维电泳分析了CK分布。在光学显微镜下使用四个没有组织学变化的下颌下腺作为标本。通过以前的改良方法提取了中间丝状蛋白，在1-2％十二烷基硫酸钠-5％2-巯基乙醇溶液中未充分提取CK。另一方面，在9.5M尿素-5％2-巯基乙醇溶液中萃取后，获得了11条分子量为40-58K的条带。尽管观察到细胞骨架蛋白的片段化，但根据均质化强度，条带没有差异。该片段化似乎是由于蛋白酶的限制性降解。免疫印迹揭示了该CK提取物与2种pan-CK抗体即识别II型碱性CK的K8.13和识别CK4，CK5，CK6，CK8，CK10，CK13，CK18的C-11的反应。在单特异性抗体中，抗CK7（LDS-68），CK8（M20），CK13（KS-1A3），CK18（CY-90）和CK19（BA17）抗体显示阳性反应。胶质纤维酸性蛋白，神经丝（NF）68 K，NF160 K或NF200 K没有反应。这些一维和二维电泳分析的结果表明正常下颌腺中存在CK5，CK7，CK8，CK13，CK14，CK15，CK17，CK18和CK19。

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