Fixation-dependent organization of core histones following DNA fluorescent in situ hybridization
Fixation-dependent organization of core histones following DNA fluorescent in situ hybridization
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DOI：
10.1007/s004120050231
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发表时间：
1997-07-01
期刊：
影响因子：
1.6
通讯作者：
BazettJones, DP
中科院分区：
文献类型：
--
作者：
Hendzel, MJ;BazettJones, DP
我们使用间接免疫荧光评估了 DNA 荧光原位杂交 (FISH) 实验中常用的不同 DNA 变性方案对染色质结构的影响。使用针对乙酰化组蛋白 H3 和 H4 的抗体表明，不同的程序在组蛋白置换程度方面存在显着差异。发现涉及多聚甲醛固定的程序与间接免疫荧光法乙酰化染色质组织的结构保存相容。这些结果为解释旨在确定单个位点染色质组织的 DNA FISH 实验提供了基础。
We have evaluated the effects of different DNA denaturation protocols commonly used in DNA fluorescent in situ hybridization (FISH) experiments on chromatin structure using indirect immunofluorescence. The use of antibodies to acetylated histones H3 and H4 demonstrates that the different procedures differ considerably in their extent of histone displacement. Procedures involving paraformaldehyde fixation were found to be compatible with the structural preservation of acetylated chromatin organization by indirect immunofluorescence. These results provide a basis for interpreting DNA FISH experiments aimed at determining chromatin organization of individual loci.