CST’s Mitotic Marker: SignalStain® Phospho-Histone H3 (Ser10) IHC Detection Kit is a "ready to use" system designed to detect the activation of histone H3 in human tissue and cell preps by immunohistochemistry. The kit utilizes the ABC immunoperoxidase method to detect endogenous levels of phosphorylated histone H3 protein. Prediluted Phospho-Histone H3 (Ser10) Antibody is bound by a biotinylated secondary antibody. Avidin DH and biotinylated horseradish peroxidase are complexed by mixing defined amounts prior to use, and the mixture subsequently binds the secondary antibody. The macromolecular complex is localized by incubation with NovaRED™ enzyme substrate.
The prediluted primary antibody, along with the ABC system, allows the user consistently to examine phosphorylated histone H3 localization and offers the highest sensitivity with the lowest background.
Mitotic Marker: SignalStain® Phospho-Histone H3 (Ser10) IHC Detection Kit detects endogenous histone H3 only when phosphorylated at serine 10. The antibody does not cross-react with other phosphorylated histones or with histone H3 modified not by phosphorylation but by acetylation. This kit was developed for and is recommended for immunohistochemistry only.
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding serine 10 of human histone H3. Antibodies are purified by protein A and peptide affinity chromatography.
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).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. SignalStain is a trademark of Cell Signaling Technology, Inc. NovaRED is a trademark of Vector Laboratories.
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