Dittmer Angela, Dittmer Jürgen
Klinik für Gynäkologie, Universität Halle, Halle, Germany.
Electrophoresis. 2006 Jul;27(14):2844-5. doi: 10.1002/elps.200500785.
Beta-actin is often used as a loading control in Western blot analysis. We analyzed the ability of beta-actin-specific antibodies to recognize differences in protein loading. We found that, at higher total protein loads as required for the detection of low-abundance proteins, beta-actin-specific antibodies failed to distinguish differences in actin protein levels. Diluting the antibody working solution or changing the incubation time had little effect on this phenomenon. This shows that beta-Actin is not a reliable loading control in Western blot analysis. In general, it appeared that, at longer incubation times, antibodies seem to be less able to pick up differences in the level of its target protein.
β-肌动蛋白常用于蛋白质免疫印迹分析中的上样对照。我们分析了β-肌动蛋白特异性抗体识别蛋白质上样差异的能力。我们发现，在检测低丰度蛋白质所需的较高总蛋白上样量时，β-肌动蛋白特异性抗体无法区分肌动蛋白蛋白水平的差异。稀释抗体工作溶液或改变孵育时间对这一现象影响不大。这表明在蛋白质免疫印迹分析中，β-肌动蛋白不是一个可靠的上样对照。总体而言，似乎在较长孵育时间下，抗体识别其靶蛋白水平差异的能力会降低。
