Product Pathways - Chromatin Regulation / Epigenetics

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Acetyl-Histone H3 Antibody Sampler Kit #9927

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
Acetyl-Histone H3 (Lys18) Antibody #9675	40 µl	W IHC-P ChIP	H M R	17	Rabbit
Histone H3 (D1H2) XP® Rabbit mAb #4499	40 µl	W IHC-P IF-IC	H M R Mk (Hm) (C) (Dm) (X) (Z) (B)	17	Rabbit IgG
Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649	40 µl	W IP IHC-P IF-IC F ChIP	H M R Mk Z Ce (Sc)	17	Rabbit IgG
Acetyl-Histone H3 (Lys14) Antibody #4318	40 µl	W IF-IC	H M R Mk (Dm) (Ce)	17	Rabbit
Acetyl-Histone H3 (Lys27) Antibody #4353	40 µl	W IP ChIP	H M R Mk (Hm) (C) (Dm) (X) (Z) (B)	17	Rabbit
Acetyl-Histone H3 (Lys56) Antibody #4243	40 µl	W IP IF-IC	H M R Mk	17	Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat

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  Ce=C. elegans
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

All antibodies in the Acetyl-Histone H3 Antibody Sampler Kit recognize histone H3 only when modified at the indicated site.

Description

The Acetyl-Histone H3 Antibody Sampler Kitprovides a fast and economical means of evaluating the acetylation sites on Histone H3. The kit contains enough primary and secondary antibodies to perform four Western mini-blot experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic acetylated peptides (KLH-coupled) corresponding to residues surrounding Lys9, Lys14, Lys18, Lys27, or Lys56 of human Histone H3. 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 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

• Allison, S.J. and Milner, J. (2003) Loss of p53 has site-specific effects on histone H3 modification, including serine 10 phosphorylation important for maintenance of ploidy. Cancer Res. 63, 6674-6679. Applications: Western Blotting
• Parsons, X. H. et al. (2003) Histone deacetylation by Sir2 generates a transcriptionally repressed nucleoprotein complex. Proc. Nat. Acad. Sci. USA 100, 1609-1614. Applications: Western Blotting
• Yoon, H. G. et al. (2003) Purification and functional characterization of the human N-CoR complex: the roles of HDAC3, TBL1 and TBLR1. EMBO J. 22, 1336-1346. Applications: Western Blotting

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

Protocols

4243:

Immunofluorescence, Immunoprecipitation, Western Blotting

4318:

Immunofluorescence, Western Blotting

4353:

ChIP Agarose, ChIP Magnetic, Immunoprecipitation, Western Blotting

4499:

IHC / Paraffin, Immunofluorescence*, Western Blotting

9649:

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

9675:

ChIP Agarose, ChIP Magnetic, IHC / Paraffin, Western Blotting

* Product-specific protocol.

Companion Products

• 9928 Histone Deacetylase (HDAC) Antibody Sampler Kit
• 2252 PRMT5/Skb1Hs Methyltransferase Antibody
• 2152 WSTF Antibody
• 7071 Phototope^®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 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

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