Overview

  • Product name

    Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade, purified
    See all Histone H3 primary antibodies
  • Description

    Rabbit polyclonal to Histone H3 (tri methyl K79) - ChIP Grade, purified
  • Host species

    Rabbit
  • Specificity

    Also expected to react with Yeast.
  • Tested applications

    Suitable for: ICC/IF, CHIPseq, WB, ChIP, Dot blotmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3 (tri methyl K79) conjugated to keyhole limpet haemocyanin.

  • Positive control

    • ICC/IF: Human osteosarcoma (U2OS) cells. ChIP: HeLaS3 cells. WB: Histone extracts from HeLa cells.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservatives: 0.05% Sodium azide, 0.05% Proclin
    Constituent: 99% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab195500 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 1/300.
CHIPseq Use at an assay dependent concentration.

1-5 μg/ChIP

WB 1/1000. Predicted molecular weight: 15 kDa.
ChIP Use at an assay dependent concentration.

1-5 μg/ChIP

Dot blot 1/20000.

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

  • 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 analysis using HeLas3 cells, labeling Histone H3 (TRI methyl K79) with ab195500. ChIP was performed using sheared chromatin from 1000,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH promoter and and for exon 2 of the inactive myoglobin gene. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K79me3 shows a preference for active promoters.

     

  • ChIP-seq analysis, labeling Histone H3 (tri methyl K79) with ab195500 at 2 µg. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The peak distribution is shown along the complete sequence and a 600 kb region of the X-chromosome (A and B) and in a 2 Mb region from chromosome 11 (C) and in a 100 kb region surrounding the GAPDH positive control gene (D).

  • Dot Blot analysis of peptides containing modified and unmodified sequences of histone H3 and H4, labeling Histone H3 (tri methyl K79) with ab195500 at 1/20000 dilution. 0.2-100 pmol of the peptide containing the respective histone modification were spotted onto the membrane.  

  • Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade, purified (ab195500) at 1/1000 dilution (in TBS-Tween containing 5% skimmed milk) + histone extracts of HeLa cells at 15 µg

    Predicted band size: 15 kDa

  • Immunofluorescent analysis of Human osteosarcoma (U2OS) cells labeling Histone H3 (tri methyl K79) with ab195500 at 1/300 dilution in blocking solution followed by row A: an anti-rabbit antibody conjugated to Alexa568 (left) or with DAPI (right), which specifically labels DNA. Rows B-C: staining of the cells with the ab195500 after incubation of the antibody with 2 ng/µl H4K20me3 and H3K79me3 peptide, respectively. Cells were fixed with 2.5% formaldehyde for 30 minutes and blocked with PBS/TX-100 containing 1% BSA.

References

This product has been referenced in:

  • Zhou R  et al. Characterization of H3 methylation in regulating oocyte development in cyprinid fish. Sci China Life Sci N/A:N/A (2018). Read more (PubMed: 30443860) »
  • Reverón-Gómez N  et al. Accurate Recycling of Parental Histones Reproduces the Histone Modification Landscape during DNA Replication. Mol Cell 72:239-249.e5 (2018). Read more (PubMed: 30146316) »
See all 2 Publications for this product

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