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Download Acetyl-Histone H4 (Lys5) Polyclonal Antibody
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Acetyl-Histone H4 (Lys5) Polyclonal Antibody Store at –20°C Catalog Number: A16719 Pub. No. MAN0009212 Rev. 1.00 Clonality: Polyclonal Amount: 100 μL Reactivity: Histone H4 Species Reactivity: Human, Mouse, Rat, Monkey Host/Class: Rabbit Product Description The nucleosome is the primary building block of chromatin, and consists of four core histone proteins (H2A, H2B, H3, and H4), which undergo multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1,2). Histone acetylation occurs mainly on the amino-terminal tail domains of histones H2A (Lys5), H2B (Lys5, 12, 15, and 20), H3 (Lys9, 14, 18, 23, 27, 36 and 56), and H4 (Lys5, 8, 12, and 16) and is important for the regulation of histone deposition, transcriptional activation, DNA replication, recombination, and DNA repair (1-3). Hyperacetylation of the histone tails neutralizes the positive charge of these domains and is believed to weaken histone-DNA and nucleosome-nucleosome interactions, thereby destabilizing chromatin structure and increasing the accessibility of DNA to various DNA-binding proteins (4,5). In addition, acetylation of specific lysine residues creates docking sites for a protein module called the bromodomain, which binds to acetylated lysine residues (6). Many transcription and chromatin regulatory proteins contain bromodomains and may be recruited to gene promoters, in part, through binding of acetylated histone tails. Histone acetylation is mediated by histone acetyltransferases (HATs), such as CBP/p300, GCN5L2, PCAF, and Tip60, which are recruited to genes by DNA-bound protein factors to facilitate transcriptional activation (3). Deacetylation, which is mediated by histone deacetylases (HDAC and sirtuin proteins), reverses the effects of acetylation and generally facilitates transcriptional repression (7,8). Product Specifications Immunogen: A synthetic peptide corresponding to the amino terminus of histone H4 in which Lys5 is acetylated Figure 1 Western blot analysis of extracts from various cell lines using Acetyl-Histone H4 (Lys5) Polyclonal Antibody. Product Applications Applications reported for this antibody include western blot, immunoprecipitation, immunohistochemistry (paraffin), immunofluorescence (immunocytochemistry), and ChIP. Because conditions may vary, it is recommended that each investigator determine the optimal amount of antibody to be used for each application. Application Western blotting Chicken, Drosophila, Xenopus, Zebrafish, Bovine, Pig, Horse, C. elegans Immunoprecipitation Apparent MW: 11 kDa Gene ID: 8359 Protein Accession No.: P62805 Lot: See product label Predicted Reactivity: Recommended Dilution 1:1000 1:25 Immunohistochemistry (paraffin) 1:1600 Immunofluorescence (immunocytochemistry) 1:800 ChIP 1:25 Storage and Handling Store reagents at –20°C. Avoid repeated freezing and thawing. Do not aliquot the antibody. For research use only. Not for use in diagnostic procedures Manufacturing Site • 7335 Executive Way • Frederick • MD 21704 • E-mail: [email protected] Stability When stored as instructed, expires one year from date of receipt unless otherwise indicated on product label. Storage Buffer 10 mM sodium HEPES (pH 7.5), 150 nM NaCl, 100 µg/mL BSA, 50% glycerol. References 1. Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51. 2. Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9. 3. Roth, S.Y. et al. (2001) Annu Rev Biochem 70, 81-120. 4. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79. 5. Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41. 6. Yang, X.J. (2004) Bioessays 26, 1076-87. 7. Haberland, M. et al. (2009) Nat Rev Genet 10, 32-42. 8. Haigis, M.C. and Sinclair, D.A. (2010) Annu Rev Pathol 5, 253-95. Figure 2 Immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A (TSA) (right), using AcetylHistone H4 (Lys5) Polyclonal Antibody (green) and a Golgin97 antibody (red). Actin filaments were labeled with a dye conjugated phalloidin (pseudocolored blue). Product Documentation To obtain a Certificate of Analysis or Safety Data Sheets (SDSs), visit www.lifetechnologies.com/support. Limited Product Warranty Life Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies’ General Terms and Conditions of Sale found on Life Technologies’ website at www.lifetechnologies.com/termsandconditions. If you have any questions, please contact Life Technologies at www.lifetechnologies.com/support. Figure 3 Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 20 μL of Acetyl-Histone H4 (Lys5) Polyclonal Antibody or 2 μL of normal rabbit IgG. The enriched DNA was quantified by real-time PCR. 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. 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The trademarks mentioned herein are the property of Life Technologies Corporation and/or its affiliates or their respective owners. For support visit www.lifetechnologies.com/support or email [email protected] www.lifetechnologies.com 28 August 2013