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Merck
  • Analysis of histone modifications in mouse neocortical neural progenitor-stem cells at various developmental stages.

Analysis of histone modifications in mouse neocortical neural progenitor-stem cells at various developmental stages.

STAR protocols (2021-09-02)
Masafumi Tsuboi, Yukiko Gotoh
摘要

Dynamic changes in histone modifications mediated by Polycomb group proteins can be indicative of the transition of gene repression mode during development. Here, we present methods for the isolation of mouse neocortical neural progenitor-stem cells (NPCs) and their culture, followed by chromatin immunoprecipitation quantitative PCR (ChIP-qPCR) techniques to examine changes in histone H2A ubiquitination patterns at various developmental stages. This protocol can be applied for both in vitro NPCs and NPCs directly isolated from mouse neocortices. For complete details on the use and execution of this protocol, please refer to (Tsuboi et al., 2018).

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Sigma-Aldrich
牛血清白蛋白 来源于牛血清, lyophilized powder, ≥96% (agarose gel electrophoresis)
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DNase I, from bovine pancreas
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白蛋白 溶液 来源于牛血清, 7.5% in DPBS, sterile-filtered, BioXtra, suitable for cell culture
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抑肽酶 来源于牛肺, saline solution, 3-7 TIU/mg protein
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胰蛋白酶抑制剂 来源于鸡蛋白, Type II-O, Partially purified ovomucoid, containing ovoinhibitor
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抗泛素组蛋白H2A抗体,克隆E6C5, clone E6C5, Upstate®, from mouse
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Rabbit Anti-Mouse IgMµ Antibody, mu chain, Upstate®, from rabbit