Comparison of Stain-Free gels with traditional immunoblot loading control methodology
Comparison of Stain-Free gels with traditional immunoblot loading control methodology
复制标题
DOI：
10.1016/j.ab.2012.08.015
复制
发表时间：
2012-11-15
影响因子：
2.9
通讯作者：
Chataway, Tim K.
中科院分区：
文献类型：
--
作者：
Colella, Alex D.;Chegenii, Nusha;Chataway, Tim K.
上样对照对于半定量Western印迹是必需的，以补偿上样误差。加载控制方法包括用针对组成型表达蛋白的抗体重新探测膜或用总蛋白染色剂染色膜。我们比较了最近发布的无蛋白质（SF）凝胶与Sypro Ruby（SR）和使用β-肌动蛋白重新探测的加载控制性能。SF凝胶表现出上级性能，因为它们更快，需要更少的步骤和消耗品。并允许在进行昂贵且耗时的蛋白质印迹之前评估电泳和蛋白质转移的质量。(C)2012 Elsevier Inc. All rights reserved.
Loading controls are necessary for semiquantitative Western blotting to compensate for loading errors. Loading control methods include the reprobing of membranes with an antibody against a constitutively expressed protein or staining the membrane with a total protein stain. We compared the loading control performance of recently released Stain-Free (SF) gels with Sypro Ruby (SR) and reprobing using beta-actin. SF gels demonstrated superior performance in that they were faster, required fewer steps and consumables. and allowed the quality of electrophoresis and Western transfer to be assessed before committing to costly and time-consuming Western blots. (C) 2012 Elsevier Inc. All rights reserved.