Hazzalin Catherine A, Mahadevan Louis C
Nuclear Signalling Laboratory, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
Methods Mol Biol. 2017;1528:173-198. doi: 10.1007/978-1-4939-6630-1_11.
Acid-urea gel electrophoresis offers significant advantages over SDS-PAGE for analysis of post-translational protein modifications, being capable of resolving proteins of similar size but varying in charge. Hence, it can be used to separate protein variants with small charge-altering differences in primary sequence, and is particularly useful in the analysis of histones whose charge variation arises from post-translational modification, such as phosphorylation or acetylation. On acid-urea gels, histones that carry multiple modifications, each with a characteristic charge, are resolved into distinct bands, the so-called "histone ladder." Thus, the extent and distribution of different modification states of histones can be visualized. Here, we describe the analysis of histone H3 by acid-urea gel electrophoresis and western blotting.
对于翻译后蛋白质修饰的分析，酸性尿素凝胶电泳相较于SDS - PAGE具有显著优势，它能够分离大小相似但电荷不同的蛋白质。因此，它可用于分离在一级序列中电荷改变差异较小的蛋白质变体，尤其在分析组蛋白时很有用，组蛋白的电荷变化源于翻译后修饰，如磷酸化或乙酰化。在酸性尿素凝胶上，带有多种修饰（每种修饰都有特定电荷）的组蛋白被分离成不同的条带，即所谓的“组蛋白梯”。因此，可以直观地看到组蛋白不同修饰状态的程度和分布。在这里，我们描述了通过酸性尿素凝胶电泳和蛋白质免疫印迹法对组蛋白H3的分析。
