RabMAb

Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

Overview

  • Product nameAnti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP GradeSee all Histone H4 primary antibodies ...
  • Description
    Rabbit monoclonal [EP1000Y] to Histone H4 (acetyl K5) - ChIP Grade
  • Tested applicationsChIP, WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis, Rice
  • Immunogen

    Synthetic peptide corresponding to residues near the N-terminus of human Histone H4.

  • Positive control
    • HeLa cells or human brain glioma. IHC-P: Human normal colon FFPE tissue sections.
  • General notes

    Produced under U.S. Patent No. 5,675,063. A 40 μl trial size is available to purchase for this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab51997 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP Use at an assay dependent concentration. See reference.
WB 1/10000 - 1/50000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
IHC-P 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/250 - 1/500.
IP 1/50.
  • Application notesIs unsuitable for Flow Cyt.
  • Target

    • FunctionCore component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
    • Sequence similaritiesBelongs to the histone H4 family.
    • Post-translational
      modifications
      Acetylation at Lys-6 (H4K5ac), Lys-9 (H4K8ac), Lys-13 (H4K12ac) and Lys-17 (H4K16ac) occurs in coding regions of the genome but not in heterochromatin.
      Citrullination at Arg-4 (H4R3ci) by PADI4 impairs methylation.
      Monomethylation and asymmetric dimethylation at Arg-4 (H4R3me1 and H4R3me2a, respectively) by PRMT1 favors acetylation at Lys-9 (H4K8ac) and Lys-13 (H4K12ac). Demethylation is performed by JMJD6. Symmetric dimethylation on Arg-4 (H4R3me2s) by the PRDM1/PRMT5 complex may play a crucial role in the germ-cell lineage.
      Monomethylated, dimethylated or trimethylated at Lys-21 (H4K20me1, H4K20me2, H4K20me3). Monomethylation is performed by SET8. Trimethylation is performed by SUV420H1 and SUV420H2 and induces gene silencing.
      Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins. Monoubiquitinated at Lys-92 of histone H4 (H4K91ub1) in response to DNA damage. The exact role of H4K91ub1 in DNA damage response is still unclear but it may function as a licensing signal for additional histone H4 post-translational modifications such as H4 Lys-21 methylation (H4K20me).
      Sumoylated, which is associated with transcriptional repression.
    • Cellular localizationNucleus. Chromosome.
    • Information by UniProt
    • Database links
      see all
    • Alternative names
      • Histone gene cluster 1, H4A antibody
      • Histone gene cluster 2, H4 antibody
      • dJ160A22.1 antibody
      • dJ160A22.2 antibody
      • dJ221C16.1 antibody
      • dJ221C16.9 antibody
      • FO108 antibody
      • H4 antibody
      • H4 histone family, member A antibody
      • H4 histone family, member B antibody
      • H4 histone family, member C antibody
      • H4 histone family, member D antibody
      • H4 histone family, member E antibody
      • H4 histone family, member G antibody
      • H4 histone family, member H antibody
      • H4 histone family, member I antibody
      • H4 histone family, member J antibody
      • H4 histone family, member K antibody
      • H4 histone family, member M antibody
      • H4 histone family, member N antibody
      • H4 histone, family 2 antibody
      • H4/A antibody
      • H4/B antibody
      • H4/C antibody
      • H4/D antibody
      • H4/E antibody
      • H4/G antibody
      • H4/H antibody
      • H4/I antibody
      • H4/J antibody
      • H4/K antibody
      • H4/M antibody
      • H4/N antibody
      • H4/O antibody
      • H4/p antibody
      • H4_HUMAN antibody
      • H4F2 antibody
      • H4F2iii antibody
      • H4F2iv antibody
      • H4FA antibody
      • H4FB antibody
      • H4FC antibody
      • H4FD antibody
      • H4FE antibody
      • H4FG antibody
      • H4FH antibody
      • H4FI antibody
      • H4FJ antibody
      • H4FK antibody
      • HIST1 cluster, H4A antibody
      • HIST1 cluster, H4B antibody
      • HIST1 cluster, H4D antibody
      • HIST2H4 antibody
      • Hist4 cluster, H4 antibody
      • HIST4H4 antibody
      • histone 1, H4a antibody
      • histone 1, H4c antibody
      • histone 1, H4d antibody
      • histone 1, H4f antibody
      • histone 1, H4h antibody
      • histone 1, H4i antibody
      • histone 1, H4j antibody
      • histone 1, H4k antibody
      • histone 1, H4l antibody
      • histone 2, H4a antibody
      • histone 2, H4b antibody
      • Histone 4 family, member M antibody
      • histone 4, H4 antibody
      • histone cluster 1, H4 antibody
      • histone cluster 1, H4a antibody
      • histone cluster 1, H4b antibody
      • histone cluster 1, H4c antibody
      • histone cluster 1, H4d antibody
      • histone cluster 1, H4e antibody
      • histone cluster 1, H4f antibody
      • histone cluster 1, H4h antibody
      • histone cluster 1, H4i antibody
      • histone cluster 1, H4j antibody
      • histone cluster 1, H4k antibody
      • histone cluster 1, H4l antibody
      • histone cluster 2, H4a antibody
      • histone cluster 2, H4b antibody
      • histone cluster 4, H4 antibody
      • Histone family, member A antibody
      • Histone family, member B antibody
      • Histone family, member D antibody
      • Histone family, member H antibody
      • Histone family, member I antibody
      • Histone family, member L antibody
      • Histone gene cluster 1, H4 antibody
      • Histone gene cluster 1, H4D antibody
      • Histone gene cluster 1, H4E antibody
      • Histone gene cluster 1, H4K antibody
      • Histone gene cluster 4, H4 antibody
      • Histone gene cluster 4, H4 histone antibody
      • Histone H4 antibody
      • histone IV, family 2 antibody
      see all

    Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade images

    • IHC image of ab51997 staining Histone H4 (acetyl K5) in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab51997, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    • All lanes : Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997) at 1/1000000 dilution

      Lane 1 : Untreated HeLa cells
      Lane 2 : TSA treated HeLa calls

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat anti-rabbit HRP labelled (1:2000)

      Predicted band size : 11 kDa
      Observed band size : 11 kDa
    • Ab51997 (1:500) staining human histone H4 on human brain glioma by immunohistochemistry using paraffin embedded tissue.Ab51997 (1:500) staining human histone H4 on human brain glioma by immunohistochemistry using paraffin embedded tissue.
    • ICC/IF image of ab51997 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51997, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    References for Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

    This product has been referenced in:
    • Kim DW  et al. A histone deacetylase inhibitor improves hypothyroidism caused by a TRa1 mutant. Hum Mol Genet 23:2651-64 (2014). Mouse . Read more (PubMed: 24381310) »
    • Ren Y  et al. Potential of adipose-derived mesenchymal stem cells and skeletal muscle-derived satellite cells for somatic cell nuclear transfer mediated transgenesis in Arbas Cashmere goats. PLoS One 9:e93583 (2014). IHC ; Goat . Read more (PubMed: 24699686) »

    See all 15 Publications for this product

    Product Wall

    We listed ChIP on the datasheet as a tested application based on a reference that used if for ChIP.

    Knutson SK et al. Liver-specific deletion of histone deacetylase 3 disrupts metabolic transcriptional networks. EMBO J 27:1017-28 (2008).
    <...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Xenopus laevis Cell (Fibroblast)
    Specification Fibroblast
    Fixative Paraformaldehyde
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    Submitted Apr 05 2013

    Thank you for contacting Abcam.

    All of our products are for research purposes only and are not certified for clinical research applications.

    I am sorry that I could not be more helpful in this matter.

    Thank you for your reply. Please keep me updated on what you decided about ab88675. I have created a discount code for you to test ab87277 in Western blot, please see below for details and let me know if you have any questions! DISCOUNT CODE: *** Expir...

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    Thanks for your call today and for letting us know about the trouble with these antibodies. As we discussed, I am sending free of charge vials of ab51997 and ab7311 on the order *** which should have arrived on November 23.    Since ab87277 and ab88675...

    Read More

    Thanks for your call today and for letting us know about the trouble with these antibodies. As we discussed, I am sending free of charge vials of ab51997 and ab7311 on the order *** which should arrive tomorrow. Since ab87277 and ab88675 have not been ...

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application Western blot
    Sample Schizosaccharomyces pombe Purified protein (Purified modified histone peptides)
    Loading amount 0.1 µg
    Specification Purified modified histone peptides
    Gel Running Conditions Non-reduced Non-Denaturing (Native)
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 03 2010

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"