Cell Signaling Technology

Product Pathways - Chromatin Regulation

Phospho-Histone H3 (Mitotic Marker) Antibody Sampler Kit #9849

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-Histone H3 (Ser10) Antibody # 9701 40 microliters W IP IHC-P IHC-F IF-IC F H M R Mk Sc C Dm Z (X) 17 Rabbit
Phospho-Histone H3 (Thr11) Antibody # 9764 40 microliters W IP IF-IC F H M R (X) 17 Rabbit
Phospho-Histone H3 (Ser28) Antibody # 9713 40 microliters W IP IF-F IF-IC F H M Hm Dm (R) (C) (B) (X) (Z) 17 Rabbit
Phospho-Histone H3 (Thr3) Antibody # 9714 40 microliters W IHC-P H M R 17 Rabbit
Histone H3 Antibody # 9715 40 microliters W IP IHC-P IF-IC H M R Mk Pg B Z (Dm) 17 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Pg=Pig  Sc=S. cerevisiae  C=Chicken  Hm=Hamster  B=Bovine  Dm=D. melanogaster  X=Xenopus  Z=Zebrafish

Specificity / Sensitivity

All antibodies in the Phospho-Histone H3 Antibody Sampler Kit recognize Histone H3 only when modified at the indicated site.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Histone H3 Antibody #9715.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with TSA (to induce histone acetylation) or serum plus calyculin A (to induce phosphorylation of H3) or both, using Phospho-Histone H3 (Ser10) Antibody #9701.

Western Blotting

Western Blotting

Western blot analysis of extracts from CHO and HeLa cells, untreated or synchronized in metaphase by treatment with 0.1 mg/ml nocodazole for 4 h, followed by isolation of metaphase cells by mitotic shake-off. Blots were probed with Phospho-Histone H3 (Ser28) Antibody #9713 (upper) or Histone H3 Antibody #9715 (lower).


Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 and HeLa cells, untreated or treated with calyculin A and 20% FCS, using Phospho-Histone H3 (Thr3) Antibody #9714 (upper) or Histone H3 Antibody #9715 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, C6 and NIH/3T3 cells, untreated or treated with nocodazole for 24 hours in the presence or absence of λ phosphatase, using Phospho-Histone H3 (Thr11) Antibody #9764 (upper) or Histone H3 Antibody #9715 (lower).

Description

The Phospho-Histone H3 (Mitotic Marker) Antibody Sampler Kit provides a fast and economical means of evaluating phosphorylation sites associated with mitosis on Histone H3. The kit contains enough primary and secondary antibodies to perform four Western blots.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Thr3, Ser10, Thr11 or Ser28 of human Histone H3. Antibodies are purified by protein A and peptide affinity chromatography.

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, 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, 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 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, 545-79.
  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.

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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