Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-Histone H3 (Ser10) Sandwich ELISA Antibody Pair #7207

Kit Includes Volume Cap Color
Total Histone H3 Capture Ab (100X) 0.4 ml Pink
Biotinylated P-Histone H3 (S 10) Detection Ab (100X) 0.4 ml Blue
HRP-Linked Streptavidin (1000X) 0.04 ml Natural

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Description

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

Specificity / Sensitivity

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.

Sandwich ELISA

Sandwich ELISA

The relationship between lysate protein concentration from untreated and serum- and Calyculin A-treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Phospho-Histone H3 (Ser10) Sandwich ELISA Antibody Pair #7207 is shown. NIH/3T3 cells were starved overnight and then given serum for 15 minutes. Calyculin A was added, the cells were incubated at 37ºC for 15 minutes and then lysed.

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, 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).

  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.

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