申请人:Nordheim, Alfred, Prof. Dr.
公开号:EP1059531A1
公开(公告)日:2000-12-13
This invention relates to methods for labelling peptides and/or proteins in preparative and analytical electrophoresis, especially as sample preparation for isoelectric focussing, wherein side chains of specific amino acid residues of the peptides and/or proteins are derivatized and labelled. The purpose of the invention is to control the isoelectric point of the peptides and/or proteins to permit isoelectric focussing of the samples with reliable results. In one embodiment of the invention cysteine residues of the peptides and/or proteins are alkylated, such that the modified amino acid side chain can be labelled, e.g. iodinated, along with tyrosine during standard conditions for tyrosine labelling. The invention permits unincorporated label to be electrophoretically removed from the sample during subsequent electrophoresis, thus dramatically improving the ratio of specific protein-bound signal to non-specific background signal upon isotopic detection.
本发明涉及在制备和分析电泳中对肽和/或蛋白质进行标记的方法,特别是作为等电点聚焦的样品制备,其中对肽和/或蛋白质的特定氨基酸残基的侧链进行衍生化和标记。本发明的目的是控制肽和/或蛋白质的等电点,以便对样品进行等电聚焦,并获得可靠的结果。在本发明的一个实施方案中,肽和/或蛋白质的半胱氨酸残基被烷基化,这样,在酪氨酸标记的标准条件下,经修饰的氨基酸侧链可与酪氨酸一起被标记,如碘化。本发明允许在随后的电泳过程中将未结合的标签从样品中电泳去除,从而大大提高了同位素检测时特异性蛋白质结合信号与非特异性背景信号的比率。