Product Pathways - Chromatin Regulation

Acetyl-Histone Antibody Sampler Kit #9933

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Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  IC=Immunocytochemistry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Pg=Pig  Y=Yeast  B=Bovine  Dr=Drosophila

Specificity / Sensitivity

Each acetyl-histone antibody recognizes only the indicated protein target modified at the indicated site. Each control histone antibody recognizes the corresponding histone regardless of its acetylation state.

Western Blotting

Western blot analysis of whole cell lysates of NIH/3T3 cells untreated, TSA-treated (to induce histone acetylation), treated with serum plus calyculin A (to induce H3 phosphorylation) or both, using Acetyl-Histone H3 (Lys9) Antibody #9671.

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 Antibody #9715.

Western Blotting

Western blot analysis of extracts from various cell lines, untreated or TSA-treated (400 nM for 12 hours), using Acetyl-Histone H2A (Lys5) Antibody #2576.

Western Blotting

Western blot analysis of extracts from various cell lines, using Histone H4 Antibody #2592.

Western Blotting

Western blot analysis of extracts from various cell lines treated with TSA (400 nM for 12 hours) or untreated, using Acetyl-Histone H4 (Lys8) Antibody #2594.

Western Blotting

Western blot analysis of extracts from various cell lines, using Histone H2B Antibody #2722.

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing nuclear localization of acetyl-histone H2A, using Acetyl-Histone H2A (Lys5) Antibody #2576.

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing nuclear localization of histone H4, using Histone H4 Antibody #2592.

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing nuclear localization of acetylated histone H4 (Lys8), using Acetyl-Histone H4 (Lys8) Antibody #2594.

IHC-P (paraffin)

Immunohistochemical staining of formalin-fixed, paraffin-embedded human breast carcinoma, showing nuclear localization of acetylated histone, using Acetyl-Histone H3 (Lys9) Antibody #9671.

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, showing nuclear localization of histone H3 , using Histone H3 Antibody #9715.

IC-ABC

Immunocytochemical staining of Vero cells untreated (left) and treated with 400 nM TSA for 12 hours (right), using Acetyl-Histone H2A (Lys5) Antibody #2576.

IC-ABC

Immunocytochemical staining of NIH/3T3 cells, using Histone H4 Antibody #2592 (left) or the same antibody preincubated with histone H4 specific peptide (right).

IC-ABC

Immunocytochemical staining of untreated cells (left) or Vero cells treated with 400 nM TSA for 12 hours (right), using Acetyl-Histone H4 (Lys8) Antibody #2594.

IC-ABC

Immunocytochemical staining of NIH/3T3 cells, untreated or TSA-treated, using Acetyl-Histone H3 (Lys9) Antibody #9671 (brown).

IF-IC

Immunocytochemical staining of NIH/3T3 cells, using Histone H3 Antibody #9715 (left) or the same antibody preincubated with specific histone H3 peptide (right).

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic acetylated peptides (KLH-coupled) corresponding to residues surrounding Lys5 of histone H2A, Lys5 of histone H2B, Lys9 of histone H3, Lys8 of histone H4, or with synthetic peptides corresponding to the amino-terminal sequences of human histone H2A, H2B, H3 or H4. Antibodies are purified by protein A and peptide affinity chromatography.

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.

Application References

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