Cell Signaling Technology

Product Pathways - Chromatin Regulation

Phospho-Histone H3 (Thr3) Antibody #9714

Applications Reactivity Sensitivity MW (kDa) Source
W IHC-P H M R Endogenous 17 Rabbit

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Histone H3 (Thr3) Antibody detects endogenous levels of histone H3 protein only when phosphorylated at Thr3. The antibody does not cross-react with other phosphorylated histones or with acetylated histones.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr3 of human histone H3. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 and HeLa cells, untreated or calyculin A and 20% FCS treated, using Phospho-Histone H3 (Thr3) Antibody (upper) or Histone H3 Antibody #9715 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing nuclear localization, using Phospho-Histone H3 (Thr3) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded MCF7 cells untreated (left) or nocodazole-treated (right), using Phospho-Histone H3 (Thr3) Antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human tonsil untreated (left) or lambda phosphatase-treated (right), using Phospho-Histone H3 (Thr3) Antibody.

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. 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.

Application References

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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