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

Overview

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

    corresponding to Human Histone H4 aa 1-100 (N terminal) (acetyl K5).
    Database link: P62805

  • Positive control
    • HeLa, NIH/3T3, C6 cells or human brain glioma, human cervical carcinoma, human normal colon FFPE, mouse liver and rat cerebral cortex tissue.
  • 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 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. PubMed: 18354499
WB 1/500000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).

For unpurified use at 1/10000- 1/50000.

IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

ICC/IF 1/5000.

For unpurified use at 1/250- 1/500.

IP 1/30.

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

  • All lanes : Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997) at 1/500000 dilution (purified)

    Lane 1 : Nuclear extract of HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 7mM Sodium Butyrate for 24 hours
    Lane 2 : Untreated nuclear extract of HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
    Lane 4 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysates
    Lane 5 : C6 (Rat glial tumor cell line) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
    Lane 6 : Untreated C6 (Rat glial tumor cell line) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

    Blocking and diluting buffer: 5% NFDM/TBST.

  • IHC image of unpurified 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.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma)treated with 500ng/m Trichostatin A for 4 hours labeling Histone H4 (acetyl K5) with purified ab51997 at 1/5000 dilution (0.1μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab195889, an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 1/200 (2.5 μg/ml). ab150077, a Goat anti rabbit IgG(Alexa Fluor® 488) secondary antibody was used at 1/1000 dilution. PBS instead of the primary antibody was used as a control. DAPI nuclear staining.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling Histone H4 (acetyl K5) with purified ab51997 at 1/500 dilution (1 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ab97051, a Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

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

    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 (purified) at 1/30 dilution (2µg) immunoprecipitating Histone H4 (acetyl K5) in HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate.

    Lane 1 (input): HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate 10ug
    Lane 2 (+): ab51997+ HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51997 in HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with TSA whole cell lysate

    For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  • ICC/IF image of unpurtified 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:
  • Zhang H  et al. Histone Modification Is Involved in Okadaic Acid (OA) Induced DNA Damage Response and G2-M Transition Arrest in Maize. PLoS One 11:e0155852 (2016). Plants . Read more (PubMed: 27196101) »
  • 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) »

See all 19 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

Submitted May 20 2015

We listed ChIP on the datasheet based on a reference that used the antibody for this application.

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

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"