| Isolating Taste Buds and Taste Cells from Vallate Papillae of C57BL/6J Mice for Detecting Transmitter Secretion
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Author:
Date:
2016-06-05
[Abstract] Mouse is a well-accepted model for studying taste bud function. Mice readily detect and respond to taste substances that humans consider to have sweet, bitter, salty, sour and umami taste qualities. A great deal of recent research on taste receptors is based on this species. Live mice are needed for these experiments because no alternative in vitro model incorporates all elements of taste transduction and peripheral signaling. The C57BL/6J strain was selected because these mice respond robustly to many taste stimuli and because of variety of transgenic animals, such as PLCβ2-GFP and GAD67-GFP, were derived from that strain. Prior analyses on behavior, nerve responses, cellular electrophysiology and molecular biology, all conducted on C57BL/6J mice will form a solid foundation for ...
[摘要] 鼠标是研究味蕾功能的一个被广泛接受的模式。小鼠容易检测和响应人们认为具有甜味，苦味，咸味，酸味和鲜味的品质的味道。最近对味觉感受器的大量研究是基于这个物种。这些实验需要活的小鼠，因为没有替代的体外模型包含所有的味觉转导和外周信号的元件。选择C57BL / 6J菌株，因为这些小鼠对许多味觉刺激反应强烈，并且由于来自该菌株的多种转基因动物，例如PLCβ2-GFP和GAD67-GFP。对C57BL / 6J小鼠进行的行为，神经反应，细胞电生理学和分子生物学的先前分析将为拟议研究奠定坚实的基础（Finger et al。，2005; Huang and Wu，2015; Huang et al。，2007 ）。因此，新鲜安乐死的动物必须用作味蕾的来源，我们将从中分离出味蕾和味道细胞。
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| Clonal Culture of Mouse Liver Progenitor Cells
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Author:
Date:
2015-10-20
[Abstract] Liver stem/progenitor cells (LPCs) are defined as bipotential cells differentiating into both hepatocytes and cholangiocytes. For analyzing their differentiation potential, clonal culture has been used for LPCs isolated by a cell sorter. In addition, we can use the culture to assess functions of target genes on differentiation potential of LPCs. This protocol describes the process of cell isolation and colony assay to examine proliferative and differentiation potential of LPCs.
[摘要] 肝干/祖细胞（LPC）被定义为分化成肝细胞和胆管细胞的双能细胞。 为了分析其分化潜力，克隆培养已经用于通过细胞分选仪分离的LPC。 此外，我们可以使用文化来评估目标基因的功能分化潜力的LPCs。 该协议描述细胞分离和集落分析的过程，以检查增殖和分化潜力的LPCs。
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| GST-tagged Yeast Protein Purification
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Author:
Date:
2011-10-05
[Abstract] Glutation S-transferase (GST) tagging is the most commonly used purification strategy for recombinant protein. It was developed with the goal of preserving the enzymatic activity by utilizing gentle elution condition of the target protein from purification matrix (Poon and Hunt., 1994). The method described here can be applied from single protein to proteome scale purification of recombinant protein from yeast (Zhu et al., 2000; Zhu et al., 2001).
[摘要] Gligation S转移酶（GST）标签是重组蛋白最常用的纯化策略。 它的开发目的是通过利用来自纯化基质的目标蛋白的温和洗脱条件来保持酶活性（Poon和Hunt，1994）。 本文所述的方法可以从单一蛋白质应用于来自酵母的重组蛋白质的蛋白质组规模纯化（Zhu et al。，2000; Zhu et al。，2001）。
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