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  • 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
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Bovine Serum Albumin solution, 7.5% in DPBS, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Trypsin inhibitor from chicken egg white, Type II-O, Partially purified ovomucoid, containing ovoinhibitor
Sigma-Aldrich
Aprotinin from bovine lung, saline solution, 3-7 TIU/mg protein
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
Roche
DNase I, from bovine pancreas