Product Pathways - Chromatin Regulation

Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 647 Conjugate) #9716

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Applications	Reactivity	Sensitivity	MW (kDa)	Source
IF-IC F	H M	Endogenous	17	Rabbit

Applications Key:  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by Western blot.

Protocols

9716:

Flow, Immunofluorescence

Specificity / Sensitivity

Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 647 Conjugate) 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. The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-6.

Description

This Cell Signaling Technology Antibody was conjugated to Alexa Fluor ® 647 fluorescent dye and tested in-house for direct Flow Cytometry in human and mouse cells. The unconjugated antibody, #9701 reacts with phospho-histone H3 (Ser10) from human, mouse, rat, and monkey. CST expects that Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 647 Conjugate) will also recognize phospho-histone H3 (Ser10) in these species.

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.

Application References

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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.