Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA.
Birmingham VA Medical Center, Birmingham, AL, USA.
Methods Mol Biol. 2021;2294:269-274. doi: 10.1007/978-1-0716-1350-4_19.
Nucleolar isolation is a crucial technique for the study of nucleolar contents and regulation of ribosome biogenesis. Lysed cells are spun through various concentrations of sucrose and magnesium chloride to separate the notoriously dense nucleoli from the rest of the cell. Here we describe isolation of nucleoli from the breast cancer cell line MDA-MB-468. The resulting nucleolar fraction is subjected to immunoblotting to confirm the purity of the nucleolar fraction.
核仁分离是研究核仁内容物和核糖体生物发生调控的关键技术。裂解细胞通过不同浓度的蔗糖和氯化镁离心，将著名的致密核仁与细胞的其他部分分离。本文描述了从乳腺癌细胞系 MDA-MB-468 中分离核仁的方法。所得核仁部分进行免疫印迹以确认核仁部分的纯度。
