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RecombinantRabMAb

Recombinant Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

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ChIP-sequencing - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • ChIP - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
  • Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP959Y] to Histone H3 (acetyl K18) - ChIP Grade
  • Suitable for: ICC/IF, ChIP, ChIP-sequencing, WB, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade
    See all Histone H3 primary antibodies

  • Description

    Rabbit monoclonal [EP959Y] to Histone H3 (acetyl K18) - ChIP Grade

  • Host species

    Rabbit

  • Specificity

    This antibody only detects Histone H3 when acetylated on Lysine 18.

  • Tested applications

    Suitable for: ICC/IF, ChIP, ChIP-sequencing, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (acetyl K18). The exact sequence is proprietary.

  • Positive control

    • WB: C6, NIH/3T3 and HeLa cells treated with 500ng/ml Trichostatin A for 4 hours. IHC-P: Human breast carcinoma and endometrium cancer tissue. Mouse liver and rat stomach tissue. ICC/IF: HeLa cells. ChIP: Chromatin prepared from HeLa cells. ChIP-seq: Chromatin prepared from HeLa 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab40888 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
ICC/IF (1)
1/1000.

For unpurified use at 1/100-1/250
ChIP
Use 5 µg for 25 µg of chromatin.
ChIP-sequencing
Use 4µg for 107 cells.
WB
1/20000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).

For unpurified use at 1/500
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
ICC/IF
1/1000.

For unpurified use at 1/100-1/250
ChIP
Use 5 µg for 25 µg of chromatin.
ChIP-sequencing
Use 4µg for 107 cells.
WB
1/20000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).

For unpurified use at 1/500
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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 H3 family.

  • Developmental stage

    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.

  • Post-translational
    modifications

    Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
    Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
    Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
    Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
    Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
    Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). 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.

  • Cellular localization

    Nucleus. Chromosome.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • H3 histone family member E pseudogene antibody
    • H3 histone family, member A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3FA antibody
    • Hist1h3a antibody
    • HIST1H3B antibody
    • HIST1H3C antibody
    • HIST1H3D antibody
    • HIST1H3E antibody
    • HIST1H3F antibody
    • HIST1H3G antibody
    • HIST1H3H antibody
    • HIST1H3I antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • histone 1, H3a antibody
    • Histone cluster 1, H3a antibody
    • Histone H3 3 pseudogene antibody
    • Histone H3.1 antibody
    • Histone H3/a antibody
    • Histone H3/b antibody
    • Histone H3/c antibody
    • Histone H3/d antibody
    • Histone H3/f antibody
    • Histone H3/h antibody
    • Histone H3/i antibody
    • Histone H3/j antibody
    • Histone H3/k antibody
    • Histone H3/l antibody
    see all

Images

  • ChIP-sequencing - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    ChIP-sequencing - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

    Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 4 µg of ab40888 [EP959Y]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.

    Additional screenshots of mapped reads can be downloaded here.

  • ChIP - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    ChIP - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

    Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10min.
    The  ChIP was performed with 25 µg of chromatin, 5 µg of  ab40888 (red), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
    Primers and probes are located in the first kb of the transcribed region.

  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/20000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for 4 hours 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: 17 kDa
    Observed band size: 17 kDa

  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/50000 dilution (Purified)

    Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) 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: 17 kDa
    Observed band size: 17 kDa

  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/50000 dilution (Purified)

    Lane 1 : C6 (Rat glial tumor glial cell) treated with 500ng/ml Trichostatin A for 4 hours whole cell lysates
    Lane 2 : C6 (Rat glial tumor glial cell) 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: 17 kDa
    Observed band size: 17 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium cancer tissue sections labeling Histone H3 with purified ab40888 at 1/4000 dilution (0.23 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) treated with 500ng/ml Trichostatin A for 4 hours cells labeling Histone H3 with purified ab40888 at 1/1000 dilution (0.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Western blot - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    All lanes : Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888) at 1/50000 dilution

    Lane 1 : C6 cell lysate, untreated.
    Lane 2 : C6 cell lysate, treated with TSA.

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 17 kDa
    Observed band size: 17 kDa
    Additional bands at: 50 kDa (possible non-specific binding), 60 kDa (possible non-specific binding)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue sections labeling Histone H3 with purified ab40888 at 1/4000 dilution (0.23 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Histone H3 with purified ab40888 at 1/4000 dilution (0.23 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    ab40888 diluted 1:100, staining acetylated histone H3 on human breast carcinoma sections.
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
    ab40888 (1/500) staining Histone H3 (acetyl K18) in asynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

    See Abreview

  • Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)
    Anti-Histone H3 (acetyl K18) antibody [EP959Y] - ChIP Grade (ab40888)

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

References (12)

Publishing research using ab40888? Please let us know so that we can cite the reference in this datasheet.

ab40888 has been referenced in 12 publications.

  • Bhaskar A  et al. Host sirtuin 2 as an immunotherapeutic target against tuberculosis. Elife 9:N/A (2020). PubMed: 32697192
  • Guo X  et al. HDAC6 is associated with the formation of aortic dissection in human. Mol Med 25:10 (2019). PubMed: 30925865
  • Cheng Z  et al. K-Ras-ERK1/2 accelerates lung cancer cell development via mediating H3K18ac through the MDM2-GCN5-SIRT7 axis. Pharm Biol 57:701-709 (2019). PubMed: 31613681
  • Xu F  et al. A combination of low-dose decitabine and chidamide resulted in synergistic effects on the proliferation and apoptosis of human myeloid leukemia cell lines. Am J Transl Res 11:7644-7655 (2019). PubMed: 31934307
  • Benoit YD  et al. Sam68 Allows Selective Targeting of Human Cancer Stem Cells. Cell Chem Biol 24:833-844.e9 (2017). PubMed: 28648376
View all Publications for this product

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (acetyl K18) antibody [EP959Y]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Kirk Mcmanus

Verified customer

Submitted Apr 12 2012

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