Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)

Overview

  • Product nameAnti-Histone H4 (acetyl K16) antibody [EPR1004]
    See all Histone H4 primary antibodies
  • Description
    Rabbit monoclonal [EPR1004] to Histone H4 (acetyl K16)
  • Specificityab109463 only detects Histone H4 acetylated on Lysine 16.
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H4 (acetyl K16).

  • Positive control
    • WB: HeLa, C6 and mouse spleen cell lysates - treated with TSA. IHC-P: Human testis, transitional cell carcinoma and colon tissues. ICC/IF: HeLa cells treated with TSA. Flow Cyt: HeLa cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109463 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
WB 1/1000 - 1/2000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
IHC-P 1/100 - 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/100 - 1/200.
Flow Cyt 1/100 - 1/200. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
  • Application notesIs unsuitable for IP.
  • 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 K16) antibody [EPR1004] images

    • All lanes : Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa cell lysates, untreated
      Lane 2 : HeLa cell lysates treated with TSA

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP-labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 11 kDa
      Observed band size : 11 kDa
    • Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified) + HeLa cell lysate - treated with TSA at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 11 kDa
      Observed band size : 11 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (purified) + HeLa cell lysate - treated with TSA at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 11 kDa
      Observed band size : 11 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified) + C6 cell lysate - treated with TSA at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 11 kDa
      Observed band size : 11 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (unpurified) + C6 cell lysate - treated with TSA at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 11 kDa
      Observed band size : 11 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (purified) + Mouse spleen tissue lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1000

      Predicted band size : 11 kDa
      Observed band size : 11 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (purified) + Mouse spleen tissue lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 11 kDa
      Observed band size : 11 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with unpurified ab109463 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with purified ab109463 at 1/150. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling Histone H4 with unpurified ab109463 at 1/100.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma labelling Histone H4 (acetly K16) with unpurified ab109463 at 1/100.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/150) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/150) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    • Overlay histogram showing HeLa cells stained with unpurified ab109463 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109463, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    • Flow cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/130. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).

    • Flow cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/200. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).

    References for Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463)

    This product has been referenced in:
    • Zheng YC  et al. 1,2,3-Triazole-Dithiocarbamate Hybrids, a Group of Novel Cell Active SIRT1 Inhibitors. Cell Physiol Biochem 38:185-93 (2016). WB ; Human . Read more (PubMed: 26784898) »
    • Mandal P  et al. H3 clipping activity of glutamate dehydrogenase is regulated by stefin B and chromatin structure. FEBS J 281:5292-308 (2014). Human . Read more (PubMed: 25263734) »

    See all 2 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (10)
    Sample Rat Cell lysate - whole cell (Primary Neurons)
    Specification Primary Neurons
    Treatment 50 nM TSA for 24h
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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    Submitted Jul 29 2013

    Application Western blot
    Sample Saccharomyces cerevisiae Cell lysate - whole cell (Whole cell lysate)
    Gel Running Conditions Reduced Denaturing (15%)
    Loading amount 60000 cells
    Specification Whole cell lysate
    Blocking step Odyssey blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 25°C
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    Submitted Aug 09 2016

    Application Immunocytochemistry/ Immunofluorescence
    Sample Fruit fly (Drosophila melanogaster) Cell (ovaries)
    Permeabilization Yes - PBST
    Specification ovaries
    Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative Formaldehyde
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    Submitted Aug 24 2015

    Application Flow Cytometry
    Sample Mouse Cell (Thymus)
    Permeabilization Yes - FIX & PERM® Cell Fixation & Cell Permeabilization Kit
    Gating Strategy Thymocytes (CD45+ Epcam- cells)
    Specification Thymus
    Fixation FIX & PERM® Cell Fixation & Cell Permeabilization Kit
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    Submitted Jun 05 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Na-citrate pH6
    Sample Mouse Tissue sections (Liver, P5)
    Specification Liver, P5
    Permeabilization Yes - Triton 0,05%
    Fixative Paraformaldehyde
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    Submitted Oct 07 2014

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