| Detection of Wnt5 in Media Conditioned by Mouse Embryonic Fibroblast
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Author:
Date:
2016-10-20
[Abstract] This protocol describes the procedure of visualizing secreted Wnt5 protein in serum free media via western blotting. This procedure can also be used to visualize other secreted proteins larger than 10,000 daltons. The work presented in this paper visualizes Wnt5 secreted by mouse embryonic fibroblast (MEF), but can be adapted to other cell lines including those transiently transfected by plasmids.
[摘要] 该协议描述了通过免疫印迹观察无血清培养基中分泌的Wnt5蛋白的过程。 该程序还可用于显现大于10,000道尔顿的其他分泌的蛋白质。 本文提出的工作可视化由小鼠胚胎成纤维细胞（MEF）分泌的Wnt5，但可以适应于其他细胞系，包括由质粒瞬时转染的那些。
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| Northwestern Blot of Protein-RNA Interaction from Young Rice Panicles
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Author:
Date:
2013-04-05
[Abstract] The northwestern assay is employed to study the interaction between protein and RNA. The RNA binding proteins tend to bind to different kinds of RNA through either known domains or unknown sequences of proteins. Rice LGD1 recombinant protein, a grass-specific novel protein with RNA binding sequences in its C-terminal, was used to probe its function as an RNA binding protein. The LGD1 comprising von Willebrand factor type-A domain (vWA), coiled-coil and nuclear localization signal (NLS) is a class of protein that localizes both in the nucleus and cytoplasm. Although LGD1 does not contain any ...
[摘要] 使用西北测定来研究蛋白质和RNA之间的相互作用。 RNA结合蛋白倾向于通过蛋白质的已知结构域或未知序列结合不同种类的RNA。将水稻LGD1重组蛋白（一种在其C末端具有RNA结合序列的草特异性新蛋白）用于探测其作为RNA结合蛋白的功能。包含von Willebrand因子A型结构域（vWA），卷曲螺旋和核定位信号（NLS）的LGD1是一类定位于细胞核和细胞质中的蛋白质。虽然LGD1不包含任何推定的RNA结合结构域，我们可以使用'RNABindR'预测软件在C末端找到高亲和力RNA结合残基（Terribilini等人，2007）。从细菌纯化的LGD1重组蛋白以某种方式甚至在变性条件下形成二聚体和单体。然而，仅二聚体形式能够结合总RNA和mRNA。由于其重现性和可靠性，我们认为该协议可以用于不同的生物。
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