#9706 Phospho-Histone H3 (Ser10) (6G3) Mouse mAb

Western Blotting

Western blot analysis of whole cell lysates from NIH/3T3 cells, untreated or treated with serum plus calyculin A (to induce phosphorylation of H3), using Phospho-Histone H3 (Ser10) (6G3) Mouse mAb (top), Phospho-Histone H3 (Ser10) Antibody #9701 (middle) or Histone H3 Antibody #9712 (bottom).

IF-F

Confocal immunofluorescent analysis of proliferating/mitotic cells labeled with Phospho-Histone H3 (Ser10) (6G3) Mouse mAb (blue) in the subventricular zone following 4 h reperfusion after cerebral ischemia. Red = EGR1 antibody #4152. Green = Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854.

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells labeled with Phospho-Histone H3 (Ser10) (6G3) Mouse mAb (red) and alpha/beta-Tubulin Antibody #2148 (green) showing different stages of the cell cycle. Nonmitotic (A), prophase (B), metaphase (C) and anaphase (D).

Flow Cytometry

Flow cytometric analysis of pacilitaxel-treated THP1 cells, using Phospho-Histone H3 (Ser10) (6G3) Mouse mAb versus propidium iodide (DNA content). The red population indicates positive Phospho-Histone H3 cells.

Flow Cytometry

Flow cytometric analysis of Phospho-Histone H3 (Ser10) (6G3) Mouse mAb staining of untreated (blue) or serum/calyculin-treated (green) Ramos cells compared to a nonspecific negative control antibody (red).

Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).

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