Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Acetylated Histone H3 Sandwich ELISA Antibody Pair #7209

Kit Includes Volume Cap Color
Total Histone H3 Capture Antibody (100X) 0.4 milliliters Pink
Acetylated-Lysine Detection Antibody (100X) 0.4 milliliters Blue
Anti-Mouse IgG, HRP-Linked Antibody (1000X) 0.04 milliliters Yellow

Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.

Species Cross-Reactivity

H M Mk

Reactivity Key:  H=Human  M=Mouse  Mk=Monkey

Description

CST's PathScan® Acetylated Histone H3 Sandwich ELISA Antibody Pair is being offered as an alternative to our PathScan® Acetylated Histone H3 Sandwich ELISA Kit #7232. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are provided for performing 4 x 96 well ELISAs. The Histone Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by an Acetylated-Lysine Detection Antibody and HRP-conjugated Anti-Mouse IgG antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450 nm is proportional to the quantity of Acetylated Histone H3 protein.*Antibodies in this kit are custom formulations specific to the kit

Sandwich ELISA

Sandwich ELISA

The relationship between lysate protein concentration from untreated and TSA treated COS cells and the absorbance at 450 nm using PathScan® Acetylated Histone H3 Sandwich ELISA Antibody Pair #7209 is shown. COS cells were lysed following overnight treatment with TSA at 37°C.

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 (2,3). 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.

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