Cell Signaling Technology

Product Pathways - Chromatin Regulation

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

Applications Reactivity MW (kDa) Source Isotype
W IP IF-F IC F H M R 17 Mouse IgG1

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-F=Immunofluorescence (Frozen)  IC=Immunocytochemistry  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Histone H3 (Ser10) (6G3) Mouse mAb detects endogenous levels of histone H3 only when phosphorylated at serine 10. The antibody does not cross-react with other phosphorylated histones or acetylated histone H3.

Source / Purification

Monoclonal antibody is produced by immunizing mice with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser10 of human histone H3.

Western Blotting

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).

IHC-FL (floating)

IHC-FL (floating)

Mitotic-specific staining of third instar wild type Drosophila larval neuroblasts, using Phospho-Histone H3 (Ser10) (6G3) Mouse mAb. Images show late prophase (A), metaphase (B) and anaphase (C, D). (provided by Marie-Louise Loupart, University of Edinburgh Scotland.)

Flow Cytometry

Flow Cytometry

Flow cytometry 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 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).

IF-IC

IF-IC

Confocal immunofluorescent images 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).

IF-F

IF-F

Confocal immunofluorescent image showing 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.


Background

Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (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, on 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 and 23 (2,3). 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 of 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 of H3 Thr3 in prophase and its dephosphorylation during anaphase (11).

  1. Workman, J.L. and Kingston, R.E. (1998) Annu. Rev. Biochem. 67, 545-579.
  2. Hansen, J.C. et al. (1998) Biochemistry 37, 17637-17641.
  3. Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-45.
  4. Cheung, P. et al. (2000) Cell 103, 263-271.
  5. Bernstein, B.E. and Schreiber, S.L. (2002) Chem. Biol. 9, 1167-1173.
  6. Jaskelioff, M. and Peterson, C.L. (2003) Nat. Cell Biol. 5, 395-399.
  7. Thorne, A.W. et al. (1990) Eur. J. Biochem. 193, 701-713.
  8. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-360.
  9. Goto, H. et al. (1999) J. Biol. Chem. 274, 25543-25549.
  10. Preuss, U. et al. (2003) Nucleic Acids Res. 31, 878-885.
  11. Dai, J. et al. (2005) Genes Dev. 19, 472-488.

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