| Phenol-based Total Protein Extraction from Lily Plant Tissues
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Author:
Date:
2015-02-20
[Abstract] The phenol-based total protein extraction method is unique in that water-soluble components such as polyphenolic compounds and nucleic acids can be easily removed. Thus, total protein is free from contaminants and allows for high quality two–dimensional gel electrophoresis. The phenol-based extraction of total protein was used in various lily organs and may likely apply to other plants whose content of polyphenolics is high (Note 1). An additional advantage of this extraction method is that nucleic acids can be easily removed and thus, avoid adverse effects of nucleic acids on protein resolution in the gel. This method is modified from that of Hurkman and Tanaka (1986).
[摘要] 酚类总蛋白提取方法的独特之处在于可以容易地除去多酚化合物和核酸等水溶性成分。 因此，总蛋白不含污染物，并允许高质量的二维凝胶电泳。 总蛋白的苯酚提取用于各种百合器官，可能适用于多酚含量高的其他植物（注1）。 该提取方法的另一个优点是，可以容易地除去核酸，从而避免核酸对凝胶中蛋白质分辨的不利影响。 这种方法从Hurkman和Tanaka（1986）的修改。
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| Separation of Microspores from Anthers of Lilium longiflorum (Lily) and Subsequent RNA Extraction
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Author:
Date:
2015-02-20
[Abstract] This protocol has been designed in order to facilitate the isolation and extraction of total RNA from microspores collected from lily anther sacs. This protocol allows the extraction of high amounts of high quality RNA, as observed in agarose gels.
[摘要] 该协议已被设计以便于从从百合花囊收集的小孢子中分离和提取总RNA。 这个协议允许大量的高质量RNA，如在琼脂糖凝胶中观察到的提取。
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| Subcellular Fractionation of Cultured Human Cell Lines
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Author:
Date:
2013-05-05
[Abstract] Subcellular localization is crucial for the proper functioning of a protein. Deregulation of subcellular localization may lead to pathological consequences and result in diseases like cancer. Immuno-fluorescent staining and subcellular fractionation can be used to determine localization of a protein. Here we discuss a protocol to separate the nuclear, cytosolic, and membrane fractions of cultured human cell lines using a centrifuge and ultracentrifuge. The membrane fraction contains plasma membranes and ER-golgi membranes, but no mitochondria or nuclear structures. The fractions can be further analyzed using Western blotting. This protocol is based on that from Dr. Richard Patten at Abcam, and was modified and utilized in a publication by Huang et al. (2012).
[摘要] 亚细胞定位对于蛋白质的正常功能是至关重要的。 亚细胞定位的失调可能导致病理结果并导致诸如癌症的疾病。 免疫荧光染色和亚细胞分级分离可用于确定蛋白质的定位。 在这里我们讨论使用离心机和超速离心机分离培养的人类细胞系的核，细胞溶质和膜部分的协议。 膜级分含有质膜和ER-高尔基体膜，但没有线粒体或核结构。 可以使用蛋白质印迹法进一步分析级分。 该协议基于来自Abcam的Richard Patten博士，并且在Huang等人的出版物中被修改和利用。 （2012）。
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