Antibody pairs have been optimized using recommended buffers, reagents, plates and the included protocol. Solutions should be made fresh daily.
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
CST's PathScan® Phospho-Histone H3 (Ser10) Sandwich ELISA Antibody Pair is being offered as an alternative to our PathScan® Phospho-Histone H3 (Ser10) Sandwich ELISA Kit #7155. Capture and Detection antibodies (100X stocks) and HRP-conjugated Streptavidin (1000X stock) are supplied. Sufficient reagents are provided for performing 4 x 96 well ELISAs. Histone Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by a Biotinylated Phospho-Histone H3 (Ser10) Detection Antibody and HRP-conjugated Streptavidin. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450 nm is proportional to the quantity of Phospho-Histone H3 (Ser10) protein.
*Antibodies in this kit are custom formulations specific to the kit
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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).
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