Protein purification and analysis: next generation Western blotting techniques.
Protein purification and analysis: next generation Western blotting techniques.
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DOI：
10.1080/14789450.2017.1388167
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发表时间：
2017-11
影响因子：
3.4
通讯作者：
Gomes AV
中科院分区：
文献类型：
--
作者：
Mishra M;Tiwari S;Gomes AV
Western blotting是分子生物学和蛋白质组学中最常用的技术之一。由于western blotting是一个多步骤的协议，变化和错误可能发生在任何步骤，降低了该技术的可靠性和可重复性。最近的报告表明，一些关键步骤，如样品制备方法，使用的一抗的数量和来源，以及所使用的归一化方法，是可重复的western blot结果的关键。本文综述了western blotting在蛋白质转移和抗体验证等不同领域的进展。本文讨论了目前最先进的免疫印迹技术，并强调了下一代免疫印迹技术与其临床相关性之间的关系。在过去的十年中，通过优化western blot方案的各个方面，在创建更敏感、自动化和先进的技术方面取得了重大进展。新的方法，如单细胞分辨率western blot，毛细管电泳，DigiWest，自动微流体western blot和微芯片电泳都已经开发出来，以减少与western blot技术相关的潜在问题。仪器的创新发展和免疫印迹灵敏度的提高为增加免疫印迹的临床意义提供了新的可能性。
Western blotting is one of the most commonly used techniques in molecular biology and proteomics. Since western blotting is a multistep protocol, variations and errors can occur at any step reducing the reliability and reproducibility of this technique. Recent reports suggest that a few key steps, such as the sample preparation method, the amount and source of primary antibody used, as well as the normalization method utilized, are critical for reproducible western blot results. In this review, improvements in different areas of western blotting, including protein transfer and antibody validation, are summarized. The review discusses the most advanced western blotting techniques available and highlights the relationship between next generation western blotting techniques and its clinical relevance. Over the last decade significant improvements have been made in creating more sensitive, automated, and advanced techniques by optimizing various aspects of the western blot protocol. New methods such as single cell-resolution western blot, capillary electrophoresis, DigiWest, automated microfluid western blotting and microchip electrophoresis have all been developed to reduce potential problems associated with the western blotting technique. Innovative developments in instrumentation and increased sensitivity for western blots offer novel possibilities for increasing the clinical implications of western blot.
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影响因子：
4.1
作者：
Bass JJ;Wilkinson DJ;Rankin D;Phillips BE;Szewczyk NJ;Smith K;Atherton PJ
通讯作者：
Atherton PJ
影响因子：
2.9
作者：
AEBERSOLD, RH;PIPES, GD;KENT, SBH
通讯作者：
KENT, SBH
影响因子：
4.4
作者：
Butler, Trina M.;Elustondo, Pia A.;MacPhee, Daniel J.
通讯作者：
MacPhee, Daniel J.
影响因子：
5.4
作者：
Bell G
通讯作者：
Bell G
影响因子：
5.3
作者：
Cui, Ziyou;Hwang, Soyun Michelle;Gomes, Aldrin V.
通讯作者：
Gomes, Aldrin V.