New England Biolabs Canada
 

Product Pathways - Chromatin Regulation / Epigenetics

Acetyl-Histone H3 (Lys9/Lys14) Antibody #9677

Item# Description List Price Web Price Qty
9677L Acetyl-Histone H3 (Lys9/Lys14) Antibody - 300 µl $1,025.00
$922.50
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9677S Acetyl-Histone H3 (Lys9/Lys14) Antibody - 100 µl $433.00
$389.70
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Source
W Human, Mouse, Rat, Monkey, C. elegans Endogenous 17 Rabbit
IP
IHC-P
IF-IC
ChIP

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP

Homology

Species predicted to react based on 100% sequence homology: Zebrafish.

Directions For Use

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Specificity / Sensitivity

Acetyl-Histone H3 (Lys9/Lys14) Antibody detects endogenous levels of histone H3 only when acetylated at lysine 9 or lysine 14 . This antibody does not cross-react with other acetylated histones.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 acetylated on lysines 9 and 14. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa and NIH/3T3 cells, treated with or without 400 nM TSA for 18 hours, using Acetyl-Histone H3 (Lys9/Lys14) Antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Acetyl-Histone H3 (Lys9/Lys14) Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of NIH/3T3 cells, untreated (left) or treated with Trichostatin A (TSA) #9950 (400 nM, overnight; left), using Acetyl-Histone H3 (Lys9/Lys14) Antibody (green). Actin filaments were labeled with Alexa Fluor® 555 Phalloidin #8953 (red).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H3 (Lys9/Lys14) Antibody or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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

  • Zhang, Z. et al. (2011) Nat Med 17, 1448-55. Applications: Chromatin IP, 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!


 

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