Quantifying Western blots: Pitfalls of densitometry
Quantifying Western blots: Pitfalls of densitometry
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DOI：
10.1002/elps.200800720
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发表时间：
2009-06-01
期刊：
影响因子：
2.9
通讯作者：
Vogel, Johannes
中科院分区：
文献类型：
--
作者：
Gassmann, Max;Grenacher, Beat;Vogel, Johannes
虽然蛋白质印迹法经常被量化，但密度测定法没有记录，似乎仅仅基于传统和猜测。根据以往的经验，100篇随机选择和系统扫描的最新论文中没有一篇提供了关于蛋白质印迹结果如何转化为统计值的足够信息。然而，这些信息的重要性，变得显而易见，从我们的血浆促红细胞生成素值的各种哺乳动物使用RIA和蛋白质印迹密度测定的相关性。应用于相同蛋白质印迹的不同常见密度测定程序揭示了这些相关性的p值范围为0.000013至0.76，反映了蛋白质印迹密度测定的科学合理基础的必要性。目前，密度测定法中缺乏任何定义，这为无法控制地获取任何期望的p值打开了大门。在这里，我们提供的数据定义了在定量Western印迹时应该考虑什么，避免什么和记录什么。
Although Western blots are frequently quantified, densitometry is not documented and appears to be based merely on traditions and guesswork. Confirming previous experience, none of 100 randomly selected and systematically scanned most recent papers provided sufficient information on how Western blot results were translated into statistical values. The importance of such information, however, becomes evident from our correlations of plasma erythropoietin values of various mammals determined using RIA and Western blot densitometry. Different common densitometry procedures applied to the identical Western blot revealed p-values of these correlations ranging from 0.000013 to 0.76 reflecting the necessity of a scientifically sound basis for densitometry of Western blots. At present, the current lack of any definitions in densitometry opens the door to uncontrollable acquisition of any desired p-value. Here we provide data that define what should be considered, what avoided and what documented when quantifying Western blots.