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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
要旨

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
ウシ血清アルブミン ウシ血清由来, lyophilized powder, ≥96% (agarose gel electrophoresis)
Roche
DNase I, from bovine pancreas
Sigma-Aldrich
ウシ血清アルブミン 溶液, 7.5% in DPBS, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
アプロチニン ウシ肺由来, saline solution, 3-7 TIU/mg protein
Sigma-Aldrich
トリプシンインヒビター ニワトリ卵白由来, Type II-O, Partially purified ovomucoid, containing ovoinhibitor
Sigma-Aldrich
Anti-ubiquityl-Histone H2A Antibody, clone E6C5, clone E6C5, Upstate®, from mouse
Sigma-Aldrich
Rabbit Anti-Mouse IgMµ Antibody, mu chain, Upstate®, from rabbit