Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Phospho-Histone H3 (Ser10) Antibody #9701

Applications Reactivity Sensitivity MW (kDa) Source
W IP IHC-P IHC-F IF-IC F H M R Mk C Dm Z Sc (X) Endogenous 17 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  C=Chicken  Dm=D. melanogaster  X=Xenopus  Z=Zebrafish  Sc=S. cerevisiae
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

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

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser10 of human histone H3. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of whole cell lysates of NIH/3T3 cells, untreated, treated with TSA (to induce histone acetylation), serum plus calyculin A (to induce phosphorylation of H3) or both, using Phospho-Histone H3 (Ser10) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of phosphorylated histone H3 in paraffin-embedded human breast carcinoma showing nuclear localization using Phospho-Histone H3 (Ser10) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of HT29 cells using Phospho-Histone H3 (Ser10) Antibody. Note the specific staining of mitotic cells.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, untreated (left) or lambda phosphatase-treated (right), using Phospho-Histone H3 (Ser10) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human tonsil using Phospho-Histone H3 (Ser10) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mouse tumor using Phospho-Histone H3 (Ser10) Antibody #9701.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Histone H3 (Ser10) Antibody in the presence of control peptide (left) or Phospho-Histone H3 (Ser10) Blocking Peptide #1000 (right).

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen H1650 xenograft, showing staining of mitotic cells using Phospho-Histone H3 (Ser10) Antibody.

IC-ABC

IC-ABC

Immunocytochemical staining of NIH/3T3 cells, nocodazole-treated (2 ug/ml) and grown in 10% serum, showing cells undergoing mitosis using Phospho-Histone (Ser10) Antibody.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated Jurkat cells, using Phospho-Histone H3 (Ser10) Antibody versus propidium iodide (DNA content). The box indicates phospho-histone H3 positive cells.

Flow Cytometry

Flow Cytometry

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

IF-IC

IF-IC

Confocal microscopic image of a mitotic HeLa cell labeled with Phospho-Histone H3 (Ser10) Antibody (red) and Survivin (6E4) Mouse mAb #2802 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Background

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).

  1. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  2. Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41.
  3. Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5.
  4. Cheung, P. et al. (2000) Cell 103, 263-71.
  5. Bernstein, B.E. and Schreiber, S.L. (2002) Chem Biol 9, 1167-73.
  6. Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
  7. Thorne, A.W. et al. (1990) Eur J Biochem 193, 701-13.
  8. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
  9. Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
  10. Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.
  11. Dai, J. et al. (2005) Genes Dev 19, 472-88.
  12. Idikio, H.A. (2006) Anticancer Res 26, 4687-94.

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