Product Pathways - Chromatin Regulation / Epigenetics

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Acetyl-Histone Antibody Sampler Kit #9933

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  C=Chicken  Dm=D. melanogaster  X=Xenopus  Z=Zebrafish  B=Bovine  Sc=S. cerevisiae  Hr=Horse  Ce=C. elegans
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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 extracts from various cell lines, untreated or TSA-treated (400 nM for 12 hours), using Acetyl-Histone H2B (Lys5) Antibody #2574.

Western Blotting

Western blot Analysis of extracts from NIH/3T3, HeLa and C6 cells using Histone H2A Antibody II #2578.

Western Blotting

Western blot analysis of various cell lines using Histone H4 (L64C1) Mouse mAb #2935.

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP^® Rabbit mAb #4499.

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H2B (V119) Antibody #8135.

Western Blotting

Western blot analysis of lysates from HeLa and NIH/3T3 cells, untreated or TSA-treated (400 nM for 18 hours) using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649.

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.

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 acetylated histone H4 (Lys8), using Acetyl-Histone H4 (Lys8) Antibody #2594.

Description

The Acetyl-Histone Antibody Sampler Kit provides a fast and economical means of evaluating the acetylation states of histones H2A, H2B, H3 and H4. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic acetylated peptides corresponding to residues surrounding Lys5 of histone H2A, Lys5 of histone H2B, Lys9 of histone H3, Lys8 of histone H4, with synthetic peptides corresponding to the amino-terminal sequences of human histone H2A, or the carboxy-terminal sequences of human histone H2B. Polyclonal Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein or the amino-terminal sequences of human histone H4 protein. Acetyl-Histone H3 (Lys9) (C5B11) monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 in which Lys9 is acetylated.

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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

2574:

IHC / Paraffin, Immunoprecipitation, Western Blotting

2576:

IHC / Paraffin, Immunoprecipitation, Western Blotting

2578:

IHC / Paraffin, Immunoprecipitation, Western Blotting

2594:

Western Blotting

2935:

IHC / Paraffin, Western Blotting

4499:

IHC / Paraffin, Immunofluorescence*, Western Blotting

8135:

Western Blotting

9649:

ChIP Agarose, ChIP Magnetic, Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

* Product-specific protocol.

Companion Products

• 2152 WSTF Antibody
• 2252 PRMT5/Skb1Hs Methyltransferase Antibody
• 9927 Acetyl-Histone H3 Antibody Sampler Kit
• 9928 Histone Deacetylase (HDAC) Antibody Sampler Kit
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

For Research Use Only. Not For Use In Diagnostic Procedures.