核仁
核仁组成区
生物
核糖体生物发生
核糖体DNA
核糖体
着丝粒
核糖体RNA
DNA
基因
核糖体蛋白
细胞生物学
分子生物学
遗传学
染色体
核糖核酸
系统发育学
核心
作者
Marjolein van Sluis,Chelly van Vuuren,Brian McStay
出处
期刊:Methods in molecular biology
日期:2016-01-01
卷期号:: 3-14
被引量:11
标识
DOI:10.1007/978-1-4939-3792-9_1
摘要
3D-immunoFISH is a valuable technique to compare the localization of DNA sequences and proteins in cells where three-dimensional structure has been preserved. As nucleoli contain a multitude of protein factors dedicated to ribosome biogenesis and form around specific chromosomal loci, 3D-immunoFISH is a particularly relevant technique for their study. In human cells, nucleoli form around transcriptionally active ribosomal gene (rDNA) arrays termed nucleolar organizer regions (NORs) positioned on the p-arms of each of the acrocentric chromosomes. Here, we provide a protocol for fixing and permeabilizing human cells grown on microscope slides such that nucleolar proteins can be visualized using antibodies and NORs visualized by DNA FISH. Antibodies against UBF recognize transcriptionally active rDNA/NORs and NOP52 antibodies provide a convenient way of visualizing the nucleolar volume. We describe a probe designed to visualize rDNA and introduce a probe comprised of NOR distal sequences, which can be used to identify or count individual NORs.
科研通智能强力驱动
Strongly Powered by AbleSci AI