An overview of technical considerations for Western blotting applications to physiological research.
An overview of technical considerations for Western blotting applications to physiological research.
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DOI：
10.1111/sms.12702
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发表时间：
2017-01
影响因子：
4.1
通讯作者：
Atherton PJ
中科院分区：
文献类型：
--
作者：
Bass JJ;Wilkinson DJ;Rankin D;Phillips BE;Szewczyk NJ;Smith K;Atherton PJ
蛋白质/免疫印迹（WB）技术的应用已经达到了科学界的多个层面，现在被认为是生理学领域的常规程序。这与骨骼肌生理学有关（即解决适应运动的机制）。事实上，包括WB数据现在被认为是许多这样的生理出版物的一个重要方面，以提供对调节过程的机制性见解。尽管这种流行，也由于WB设备的无处不在和相对便宜的可用性，WB在出版物和随后的分析和解释的数据的质量可以是可变的，可能导致虚假的结论。这可能是由于实验室技术差和/或对WB中涉及的关键步骤缺乏理解，以及应该采取什么样的质量控制程序来确保可靠的数据生成。本次审查的目的是提供一个详细的描述和WB程序和技术细节的批评，从样品收集，通过准备，印迹和检测收集的数据分析。我们的目标是为读者提供更好的专业知识，以批判性地进行，评估和排除WB过程中的故障，以产生可重现和可靠的印迹。
The applications of Western/immuno-blotting (WB) techniques have reached multiple layers of the scientific community and are now considered routine procedures in the field of physiology. This is none more so than in relation to skeletal muscle physiology (i.e. resolving the mechanisms underpinning adaptations to exercise). Indeed, the inclusion of WB data is now considered an essential aspect of many such physiological publications to provide mechanistic insight into regulatory processes. Despite this popularity, also due to the ubiquitous and relatively inexpensive availability of WB equipment, the quality of WB in publications and subsequent analysis and interpretation of the data can be variable, perhaps resulting in spurious conclusions. This may be due to poor laboratory technique and/or lack of comprehension of the critical steps involved in WB and what quality control procedures should be in place to ensure robust data generation. The present review aims to provide a detailed description and critique of WB procedures and technicalities, from sample collection through preparation, blotting and detection to analysis of the data collected. We aim to provide the reader with improved expertise to critically conduct, evaluate and troubleshoot the WB process, to produce reproducible and reliable blots.
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影响因子：
4.3
作者：
Chang, Bin;Liu, Guangzhi;Liu, Jinsong
通讯作者：
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影响因子：
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DOI：
10.1152/ajpendo.00541.2012
发表时间：
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影响因子：
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作者：
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通讯作者：
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