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  1. Link

    histone-h4-acetyl-k5-antibody-ep1000y-chip-grade-ab51997.pdf

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Epigenetics and Nuclear Signaling Histones H4 Acetylated
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RecombinantRabMAb

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

  • Datasheet
Reviews (9)Q&A (1)References (82)

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ChIP - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Western blot - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • ELISA - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • ELISA - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • ELISA - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Western blot - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Immunoprecipitation - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
  • Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1000Y] to Histone H4 (acetyl K5) - ChIP Grade
  • Suitable for: ChIP, ELISA, WB, IHC-P, ICC/IF, IP
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

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Biotin Carrier Free

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Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • Product name

    Anti-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
  • Host species

    Rabbit
  • Specificity

    In addition to H4K5Ac, this antibody also detects H4K8Ac (Histone H4 acetylated on Lysine 8) at high antigen coating concentration.
  • Tested applications

    Suitable for: ChIP, ELISA, WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Recombinant fragment
    Predicted to work with: Xenopus laevis, Rice
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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. ChIP: Chromatin was prepared from MEF cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1000Y
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Histones
    • H4
    • Acetylated
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Biotin Anti-Histone H4 (acetyl K5) antibody [EP1000Y] (ab202886)
    • Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - BSA and Azide free (ab232507)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • NIH/3T3 whole cell lysate (ab7179)
  • Recombinant Protein

    • Recombinant Human Histone H4 protein (ab198115)
  • Related Products

    • Human Histone H4 peptide (ab13843)
    • Human Histone H4 (symmetric di methyl R3) peptide (ab14791)
    • Human Histone H4 (tri methyl K20) peptide (ab17567)

Applications

The Abpromise guarantee

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 (3)
Use 5 µg for 25 µg of chromatin.
ELISA
Use at an assay dependent concentration.
WB (2)
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 IHC antigen retrieval protocols.

ICC/IF (2)
1/5000.

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

IP
1/30.
Notes
ChIP
Use 5 µg for 25 µg of chromatin.
ELISA
Use at an assay dependent concentration.
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 IHC antigen retrieval protocols.

ICC/IF
1/5000.

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

IP
1/30.

Target

  • Function

    Core 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 similarities

    Belongs 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 localization

    Nucleus. Chromosome.
  • Target information above from: UniProt accession P62805 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 121504 Human
    • Entrez Gene: 554313 Human
    • Entrez Gene: 8294 Human
    • Entrez Gene: 8359 Human
    • Entrez Gene: 8360 Human
    • Entrez Gene: 8361 Human
    • Entrez Gene: 8362 Human
    • Entrez Gene: 8363 Human
    • Entrez Gene: 8364 Human
    • Entrez Gene: 8365 Human
    • Entrez Gene: 8366 Human
    • Entrez Gene: 8367 Human
    • Entrez Gene: 8368 Human
    • Entrez Gene: 8370 Human
    • Entrez Gene: 100041230 Mouse
    • Entrez Gene: 319155 Mouse
    • Entrez Gene: 319156 Mouse
    • Entrez Gene: 319157 Mouse
    • Entrez Gene: 319158 Mouse
    • Entrez Gene: 319159 Mouse
    • Entrez Gene: 319160 Mouse
    • Entrez Gene: 319161 Mouse
    • Entrez Gene: 320332 Mouse
    • Entrez Gene: 326619 Mouse
    • Entrez Gene: 326620 Mouse
    • Entrez Gene: 674678 Mouse
    • Entrez Gene: 69386 Mouse
    • Entrez Gene: 97122 Mouse
    • Entrez Gene: 64627 Rat
    • Entrez Gene: 447787 Xenopus laevis
    • Omim: 142750 Human
    • Omim: 602822 Human
    • Omim: 602823 Human
    • Omim: 602824 Human
    • Omim: 602825 Human
    • Omim: 602826 Human
    • Omim: 602827 Human
    • Omim: 602828 Human
    • Omim: 602829 Human
    • Omim: 602830 Human
    • Omim: 602831 Human
    • Omim: 602833 Human
    • SwissProt: P62805 Human
    • SwissProt: P62806 Mouse
    • SwissProt: P62804 Rat
    • SwissProt: P62799 Xenopus laevis
    • Unigene: 143080 Human
    • Unigene: 247816 Human
    • Unigene: 248172 Human
    • Unigene: 248178 Human
    • Unigene: 248179 Human
    • Unigene: 278483 Human
    • Unigene: 46423 Human
    • Unigene: 528055 Human
    • Unigene: 533295 Human
    • Unigene: 55468 Human
    • Unigene: 591790 Human
    • Unigene: 655235 Human
    • Unigene: 662174 Human
    • Unigene: 706635 Human
    • Unigene: 718935 Human
    • Unigene: 144300 Mouse
    • Unigene: 14775 Mouse
    • Unigene: 158272 Mouse
    • Unigene: 227295 Mouse
    • Unigene: 228709 Mouse
    • Unigene: 246720 Mouse
    • Unigene: 255646 Mouse
    • Unigene: 260530 Mouse
    • Unigene: 261642 Mouse
    • Unigene: 261662 Mouse
    • Unigene: 261664 Mouse
    • Unigene: 377875 Mouse
    • Unigene: 442307 Mouse
    • Unigene: 85592 Xenopus laevis
    see all
  • Alternative names

    • dJ160A22.1 antibody
    • dJ160A22.2 antibody
    • dJ221C16.1 antibody
    • dJ221C16.9 antibody
    • FO108 antibody
    • H4 antibody
    • H4.k 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/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
    • H4FM antibody
    • H4FN antibody
    • H4M antibody
    • HIST1H4A antibody
    • HIST1H4B antibody
    • HIST1H4C antibody
    • HIST1H4D antibody
    • HIST1H4E antibody
    • HIST1H4F antibody
    • HIST1H4H antibody
    • HIST1H4I antibody
    • HIST1H4J antibody
    • HIST1H4K antibody
    • HIST1H4L antibody
    • HIST2H4 antibody
    • HIST2H4A antibody
    • Hist4h4 antibody
    • Histone 1 H4a antibody
    • Histone 1 H4b antibody
    • Histone 1 H4c antibody
    • Histone 1 H4d antibody
    • Histone 1 H4e 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 4 H4 antibody
    • Histone H4 antibody
    • MGC24116 antibody
    see all

Images

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

    Chromatin was prepared from MEF (Mouse embryonic fibroblast cell line) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab51997 (red), and 20 μl protein A/G sepharose beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Western blot - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    Western blot - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    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
    All lanes : 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

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

    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) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

    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.

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

    Direct ELISA with Histone H4 K5ac peptide, Histone H4 K8ac peptide, Histone H4 K16me1 peptide, Histone H4 K16me3 peptide, and Histone H4 unmodified peptide, all at 10ng/ml. ab51997 used as the primary antibody at a range of 0~1000ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) used as the secondary antibody at 1:2500 dilution.

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

    Direct ELISA with Histone H4 K5ac peptide, Histone H4 K8ac peptide, Histone H4 K16me1 peptide, Histone H4 K16me3 peptide, and Histone H4 unmodified peptide, all at 100ng/ml. ab51997 used as the primary antibody at a range of 0~1000ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) used as the secondary antibody at 1:2500 dilution.

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

    Direct ELISA with Histone H4 K5ac peptide, Histone H4 K8ac peptide, Histone H4 K16me1 peptide, Histone H4 K16me3 peptide, and Histone H4 unmodified peptide, all at 1000ng/ml. ab51997 used as the primary antibody at a range of 0~1000ng/ml. Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) used as the secondary antibody at 1:2500 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

    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) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

    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.

  • Western blot - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    Western blot - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    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
    All lanes : Goat anti-rabbit HRP labelled (1:2000)

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

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

    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 for detection (1/10000).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade (ab51997)

    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.

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

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (82)

Publishing research using ab51997? Please let us know so that we can cite the reference in this datasheet.

ab51997 has been referenced in 82 publications.

  • Shalini V  et al. Genome-wide occupancy reveals the localization of H1T2 (H1fnt) to repeat regions and a subset of transcriptionally active chromatin domains in rat spermatids. Epigenetics Chromatin 14:3 (2021). PubMed: 33407810
  • Cai Q  et al. A fungal sirtuin modulates development and virulence in the insect pathogen, Beauveria bassiana. Environ Microbiol N/A:N/A (2021). PubMed: 33817929
  • Lin Y  et al. Environmental enrichment implies GAT-1 as a potential therapeutic target for stroke recovery. Theranostics 11:3760-3780 (2021). PubMed: 33664860
  • Li TY  et al. The transcriptional coactivator CBP/p300 is an evolutionarily conserved node that promotes longevity in response to mitochondrial stress. Nat Aging 1:165-178 (2021). PubMed: 33718883
  • Jian Y  et al. Actin-like protein 6A/MYC/CDK2 axis confers high proliferative activity in triple-negative breast cancer. J Exp Clin Cancer Res 40:56 (2021). PubMed: 33541412
View all Publications for this product

Customer reviews and Q&As

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1-10 of 10 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Colorectal carcinoma)
Permeabilization
No
Specification
Colorectal carcinoma
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Acetone
Read More

Abcam user community

Verified customer

Submitted Dec 23 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (CRC cell line)
Permeabilization
No
Specification
CRC cell line
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: RT°C
Fixative
Acetone
Read More

Abcam user community

Verified customer

Submitted Oct 01 2021

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

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (MCF7 breast cancer cell line)
Specification
MCF7 breast cancer cell line
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Read More

DR. Sankari Nagarajan

Verified customer

Submitted Apr 12 2019

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

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Arabidopsis thaliana Tissue lysate - nuclear (complete seedlings (10 days old))
Negative control
IgG (included in the Abcam kit)
Specification
complete seedlings (10 days old)
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Positive control
actin
Read More

Abcam user community

Verified customer

Submitted Dec 05 2018

Western blot abreview for Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (THP-1)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
THP-1
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Aliaksandra Radzisheuskaya

Verified customer

Submitted Oct 17 2018

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

Excellent
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - whole cell (THP-1 cells)
Negative control
no antibody control, gene desert region
Specification
THP-1 cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Positive control
active gene promoter
Read More

Aliaksandra Radzisheuskaya

Verified customer

Submitted Oct 17 2018

Flow Cytometry abreview for Anti-Histone H4 (acetyl K5) antibody [EP1000Y] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted May 20 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H4 (acetylated) (acetyl K5) antibody [EP1000Y] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Apr 05 2013

Western blot abreview for Anti-Histone H4 (acetylated) (acetyl K5) antibody [EP1000Y] - ChIP Grade

Below Average
Abreviews
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Feb 03 2010

Question

Do you have any recommendation for the concentration of this antibody (H4K5ac) per ChIP, so I decide how much to buy?

Read More

Abcam community

Verified customer

Asked on Dec 02 2013

Answer

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).

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2323257/

The authors do not give much detail of their procedure except this:

The ChIP assay was performed following the standard protocol of the Chromatin Immunoprecipitation Assay Kit from Upstate Cell Signaling, with slight modifications using minced tissue for formaldehyde crosslinking. DNA was purified using the Qiagen PCR Purification Kit. Primers designed to the designated promoter sequences were used for quantification of histone acetylation using Q-PCR, with histone H3 and input sonicated DNA as reference controls.

I suggest testing a range of amounts. Our general recommendation is 1-10ug of antibody per 25ug of DNA. Here is a link to our general cross-linked ChIP protocol:

https://www.abcam.com/index.html?pageconfig=resource&rid=11698.

We also have a ChIP kit of our own, ab500.

Read More

Tom Ruyle

Abcam Scientific Support

Answered on Dec 02 2013

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