The AMeX method: a multipurpose tissue-processing and paraffin-embedding method. Extraction of protein and application to immunoblotting.
The AMeX method: a multipurpose tissue-processing and paraffin-embedding method. Extraction of protein and application to immunoblotting.
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AMeX 方法：一种多用途组织处理和石蜡包埋方法。
DOI：
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发表时间：
1992
影响因子：
6
通讯作者：
Setsuo Hirohashi
中科院分区：
文献类型：
--
作者：
Yuichi Sato;Kiyoshi Mukai;S. Furuya;Toru Kameya;Setsuo Hirohashi
作者先前报道了一种新的固定和石蜡包埋方法（AMeX方法），该方法保留了许多抗原以及通常被常规福尔马林固定和石蜡包埋过程破坏的高分子量DNA和RNA。在本研究中，作者分析了十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹法固定的组织中的蛋白质的保存。脱蜡后从经AMeX处理的组织切片中提取蛋白质的方法与从新鲜组织中提取的方法相同。从50 mg（湿重）AMeX处理的小鼠肝组织中提取的蛋白质总量与新鲜组织相同。SDS-聚丙烯酰胺凝胶上蛋白质的电泳迁移率和染色强度，以及用几种抗体的免疫印迹图案和染色强度，对于AMeX处理的和新鲜的组织是相同的。在石蜡块中储存2年，蛋白质降解最小。作者还表明，培养细胞的沉淀可以通过这种方法进行免疫分析。这种新的固定和石蜡包埋方法是一个有用的工具，获得信息的形态特征和免疫化学和分子生物学数据之间的相关性。
The authors have previously reported a new fixation and paraffin-embedding method (the AMeX method), which preserves many antigens as well as high molecular-weight DNA and RNA that are normally destroyed by the routine formalin fixation and paraffin-embedding process. In the present study, the authors analyzed the preservation of protein suitable for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting in tissue fixed by the AMeX method. The method used for extraction of protein from AMeX-processed tissue sections after deparaffinization was the same as that for extraction from fresh tissues. The total amount of protein extracted from 50-mg (wet weight) AMeX-processed mouse liver tissue was the same as that from fresh tissue. The electrophoretic mobility and staining intensity of protein on SDS-polyacrylamide gel, and the immunoblotting pattern and staining intensity with several antibodies, were identical for both AMeX-processed and fresh tissue. Degradation of protein was minimal for storage periods of 2 years in paraffin block. The authors also showed that pellets of cultured cells can be processed by this method for immunologic analysis. This new fixation and paraffin-embedding method is a useful tool for obtaining information on correlations between morphologic features and immunochemical and molecular biological data.
对从保存在标准组织学固定剂中的组织中提取的角蛋白多肽进行免疫印迹。
DOI：
10.1177/34.5.2422250
发表时间：
1986
期刊：
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
影响因子：
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作者：
Clark,RK;Damjanov,I
通讯作者：
Damjanov,I