Phos-tag SDS-PAGE データ集 Histone H3

Histone H3

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Mw; 15,400

泳動結果

HeLa 細胞を，Ser/Thr キナーゼ阻害剤のcalyclinA(100 nM, 30 min)処理後
細胞抽出液をPhos-tag gel で泳動し
Histone H3のリン酸化亢進を調べています。

calyclinAで処理すると，リン酸化型バンドが３本観察されます。
それぞれを，部位特異的抗リン酸化抗体との交差性から，
右に示したように同定しました。

Reference	Kinoshita, E., Kinoshita-Kikuta, E., Koike, T. Phos-tag SDS-PAGE systems for phosphorylation　profiling of proteins with a wide range of molecular　masses under neutral pH conditions *Proteomics*, 12, 192-202 (2012) 10.1002/pmic.201100524
ゲルの条件	Zn²⁺–Phos-tag:　25µM Polyacrylamide: 12%(w/v)
検出方法	WB: anti-HistoneH3 anti-pHistoneH3(pT3) anti-pHistoneH3(pS10) anti-pHistoneH3(pT11) anti-pHistoneH3(pS28)

Reference

Kinoshita, E., Kinoshita-Kikuta, E., Koike, T. Phos-tag SDS-PAGE systems for phosphorylation　profiling of proteins with a wide range of molecular　masses under neutral pH conditions Proteomics, 12, 192-202 (2012)

10.1002/pmic.201100524

ゲルの条件

Zn²⁺–Phos-tag:　25µM

Polyacrylamide: 12%(w/v)

検出方法

　WB:

anti-HistoneH3
anti-pHistoneH3(pT3)
anti-pHistoneH3(pS10)
anti-pHistoneH3(pT11)
anti-pHistoneH3(pS28)