管家蛋白:在神经元分化过程中作为参考的局限性

Z. Castaño, R. Kypta
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引用次数: 13

摘要

在western blots中使用“管家蛋白”作为加载对照,要求它们不改变其表达水平。然而,已经观察到偏离这一规则的情况。本研究采用western blot方法研究了维甲酸诱导的胚胎癌NTera-2和神经母细胞瘤SH-SY5Y细胞分化过程中常用蛋白HSP60、-微管蛋白、-肌动蛋白和GAPDH的表达。-actin是一个较差的负载控制,在分化过程中显著增加,而HSP60和GAPDH在这些分析中表达更均匀。我们的结果强调了在这种生物学背景下选择准确的负载控制的重要性。
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Housekeeping Proteins: Limitations as References During Neuronal Differentiation
The use of "Housekeeping proteins" as loading controls in western blots requires that they do not change their level of expression. However, deviations from this rule have been observed. This study investigated the expression of the commonly used proteins HSP60, -tubulin,  -actin and GAPDH by western blot during retinoic acid-induced differentia- tion of the embryonal carcinoma NTera-2 and the neuroblastoma SH-SY5Y cell lines. -actin was found to be a poor loading control, increasing significantly during differentiation, whereas HSP60 and GAPDH were the more evenly ex- pressed of those analysed. Our results underline the importance of selecting accurate loading controls in this biological context.
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