Product Pathways - Chromatin Regulation

Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 488 Conjugate) #9708

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

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

Specificity / Sensitivity

Phospho-Histone H3 (Ser10) Antibody (Alexa Fluor® 488 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 rabbits with a synthetic phospho-peptide (KLH-coupled) 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®488 under optimal conditions with an F/P ratio of 2-6.

Description

This Cell Signaling Technology Antibody was conjugated to Alexa Fluor ® 488 fluorescent dye and tested in-house for direct Flow Cytometry and Immunofuorescence 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® 488 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 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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Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

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.