Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting
Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting
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DOI：
10.1016/j.ab.2014.08.027
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发表时间：
2014-12-15
影响因子：
2.9
通讯作者：
de Medina, F. Sanchez
中科院分区：
文献类型：
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作者：
Rivero-Gutierrez, B.;Anzola, A.;de Medina, F. Sanchez
目前的常规做法是要求在蛋白质印迹中测量管家参考，包括肌动蛋白、甘油醛-3-磷酸脱氢酶、β-微管蛋白等，因为这是RNA印迹中的规则。可逆丽春红染色已成功地应用于检查凝胶的相等加载。在这里，我们测试了一种新的技术，从Bio-Rad的无酶凝胶，对丽春红染色和管家蛋白免疫检测在不同的条件下。我们的研究结果表明，无酶凝胶优于丽春红染色，两者都比管家蛋白作为上样对照更一致。(C)2014作者爱思唯尔公司出版
It is currently a routine practice to require a measurement of a housekeeping reference, including actin, glyceraldehyde-3-phosphate dehydrogenase, beta-tubulin, among others, in Western blots, as it is the rule in RNA blots. Reversible Ponceau staining has been applied successfully to check equal loading of gels. Here we test a new technique, with the Stain-Free gels from Bio-Rad, against both Ponceau staining and housekeeping protein immunodetection under different conditions. Our results show that Stain-Free gels outperform Ponceau staining and that both are more consistent than housekeeping proteins as a loading control. (C) 2014 The Authors. Published by Elsevier Inc.