用于测量蛋白质降解的脉冲追踪分析。
Pulse-chase analysis to measure protein degradation.
作者信息
Simon Einav, Kornitzer Daniel
机构信息
Faculty of Medicine, Technion - I.I.T. and Rappaport Institute for Biomedical Research, Haifa, Israel.
Faculty of Medicine, Technion - I.I.T. and Rappaport Institute for Biomedical Research, Haifa, Israel.
出版信息
Methods Enzymol. 2014;536:65-75. doi: 10.1016/B978-0-12-420070-8.00006-4.
In order to measure protein half-life by pulse-chase analysis, cells are incubated with labeled precursors, [(35)S]-methionine and [(35)S]-cysteine, for a short time (pulse), after which they are washed from the labeled compound and incubated with excess cold precursor (chase). The decay of the amount of protein labeled during the pulse period is measured during the chase with cold precursor.
为了通过脉冲追踪分析来测量蛋白质半衰期，细胞先与标记的前体物质[³⁵S] - 甲硫氨酸和[³⁵S] - 半胱氨酸一起孵育一段短时间（脉冲），之后将标记化合物洗去，再与过量的冷前体物质一起孵育（追踪）。在与冷前体物质的追踪过程中，测量脉冲期标记的蛋白质数量的衰减情况。
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