Multiplexed isobaric tagging protocols for quantitative mass spectrometry approaches to auditory research.
Multiplexed isobaric tagging protocols for quantitative mass spectrometry approaches to auditory research.
复制标题
用于听觉研究定量质谱方法的多重同量异位标记协议。
DOI：
10.1007/978-1-59745-523-7_21
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Turcan,Sevin
中科院分区：
文献类型：
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作者：
Vetter,DouglasE;Basappa,Johnvesly;Turcan,Sevin
现代生物学家有大量的技术可以用来评估样品中任何感兴趣的分子的存在和相对或绝对量。然而，实施这些程序中的大多数可能是一项艰巨的任务，即使是在有经验的研究人员的实验室里也是如此。仅仅选择要遵循的方案就可能花费数周时间，同时检查所有细微差别，并确定方案是否将(A)给出期望的结果，(B)产生可解释和无偏见的数据，以及(C)服从感兴趣的样本。我们在这里详细介绍了一种稳健的程序，用于标记复杂裂解液中的蛋白质，以便在实验操作后最终对蛋白质丰度进行差异量化。在标记程序取得成功结果后，样品将被提交进行质谱分析，从而进行多肽定量和蛋白质鉴定。虽然我们将专注于培养细胞，但我们将指出可用于标记从其他组织产生的裂解产物的程序，以及此类样本所需的任何微小修改。我们还将概述(但不是完全记录)我们实验室在质谱分析之前使用的其他标记蛋白质的策略，并描述在哪些条件下每个过程可能是可取的。这一章没有涵盖的是对质谱学(仪器、理论等)的最简单的介绍，我们也不试图以用于后曲棍球分析的软件的方式来涵盖太多。这两个主题取决于一个人的资源，如果适用，还取决于一个人的合作者。我们强烈建议读者就本文未涉及的主题寻求专家建议。
Modern biologists have at their disposal a large array of techniques used to assess the existence and relative or absolute quantity of any molecule of interest in a sample. However, implementing most of these procedures can be a daunting task for the first time, even in a lab with experienced researchers. Just choosing a protocol to follow can take weeks while all of the nuances are examined and it is determined whether a protocol will (a) give the desired results, (b) result in interpretable and unbiased data, and (c) be amenable to the sample of interest. We detail here a robust procedure for labeling proteins in a complex lysate for the ultimate differential quantification of protein abundance following experimental manipulations. Following a successful outcome of the labeling procedure, the sample is submitted for mass spectrometric analysis, resulting in peptide quantification and protein identification. While we will concentrate on cells in culture, we will point out procedures that can be used for labeling lysates generated from other tissues, along with any minor modifications required for such samples. We will also outline, but not fully document, other strategies used in our lab to label proteins prior to mass spectrometric analysis, and describe under which conditions each procedure may be desirable. What is not covered in this chapter is anything but the most brief introduction to mass spectrometry (instrumentation, theory, etc.), nor do we attempt to cover much in the way of software used forpost hocanalysis. These two topics are dependant upon one’s resources, and where applicable, one’s collaborators. We strongly encourage the reader to seek out expert advice on topics not covered here.
基于结构的蛋白质-蛋白质相互作用的预测。
影响因子：
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作者：
G. Fernández;L. Serrano
通讯作者：
L. Serrano
大鼠脑突触连接蛋白制剂的蛋白质成分。
DOI：
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发表时间：
1996
期刊：
Brain Research. Molecular Brain Research
影响因子：
--
作者：
K. Langnaese;C. Seidenbecher;H. Wex;B. Seidel;K. Hartung;U. Appeltauer;A. Garner;B. Voss;B. Mueller;C. Garner;E. D. Gundelfinger
通讯作者：
E. D. Gundelfinger