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Merck

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

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

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
Przeciwciało anty-ubikwitylo-histonowe H2A, klon E6C5, clone E6C5, Upstate®, from mouse
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥96% (agarose gel electrophoresis)
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Bovine Serum Albumin solution, 7.5% in DPBS, sterile-filtered, BioXtra, suitable for cell culture
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Aprotinin from bovine lung, saline solution, 3-7 TIU/mg protein
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Trypsin inhibitor from chicken egg white, Type II-O, Partially purified ovomucoid, containing ovoinhibitor
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DNaza I, from bovine pancreas
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Królicze przeciwciało anty-Mysie IgMµ, łańcuch mu, Upstate®, from rabbit