Overview

  • Product name
  • Description
    Rabbit polyclonal to Histone H3
  • Specificity
    We have data to suggest that this antibody can detect unmodified histone H3 and acetylated K9 histone H3.
  • Tested applications
    Suitable for: ICC/IF, ELISA, IHC-P, WB, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3.
    Database link: P68431
    (Peptide available as ab154693)

  • Positive control
    • Human lung carcinoma tissue and RAW264.7 cell extracts treated with TSA (400nM, hours).

Properties

Applications

Our Abpromise guarantee covers the use of ab61251 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 1/50.
ELISA 1/5000.
IHC-P 1/50 - 1/100.
WB 1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
ChIP Use at an assay dependent concentration. PubMed: 24672748

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 A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3FA antibody
    • Hist1h3a antibody
    • HIST1H3B antibody
    • HIST1H3C antibody
    • HIST1H3D antibody
    • HIST1H3E antibody
    • HIST1H3F antibody
    • HIST1H3G antibody
    • HIST1H3H antibody
    • HIST1H3I antibody
    • HIST1H3J antibody
    • histone 1, H3a antibody
    • Histone cluster 1, H3a 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

  • ab61251 at 1/50 - 1/100 dilution staining Histone H3 in human lung carcinoma by Immunohistochemistry, Paraffin-embedded tissue, in the absence or presence of the immunising peptide.
  • All lanes : Anti-Histone H3 antibody (ab61251) at 1/500 dilution

    Lane 1 : RAW264.7 cell extracts treated with TSA (400nM, hours)
    Lane 2 : RAW264.7 cell extracts treated with TSA (400nM, hours) with the immunising peptide


    Predicted band size : 15 kDa
    Observed band size : 15 kDa
  • ab61251 staining Histone H3 in a human glioblastoma cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 (0.1%) in PBS and blocked with 0.5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/50 in PBS + 0.5% BSA) for 16 hours at 4°C. An FITC-conjugated Goat anti-rabbit monoclonal (1/80) was used as the secondary antibody. Nuclei were counterstained with Hoechst (blue).

    See Abreview

  • ab61251 staining Histone H3 in Rat choroid plexus cells by Immunocytochemistry/ Immunofluorescence. Cells were PFA-fixed and permeabilized in 0.1% Triton X-100 in PBS prior to blocking in 0.5% BSA in TBS-Tween for 20 minutes at room temperature. The primary antibody was diluted 1/50 in 0.5% BSA/PBS and incubated with the sample for 16 hours at 4°C. The secondary antibody was TRITC-conjugated goat anti-rabbit polyclonal, diluted 1/400. Nuclei were counterstained with Hoechst.

    See Abreview

  • ELISA data for Anti-Histone H3 antibody, ab61251, which shows that the antibody at various dilutions detects both non-acetylated (black line) and acetylated (red line) histone H3.

References

This product has been referenced in:
  • Yang S  et al. Probucol ameliorates renal injury in diabetic nephropathy by inhibiting the expression of the redox enzyme p66Shc. Redox Biol 13:482-497 (2017). Read more (PubMed: 28728079) »
  • Xiong Y  et al. Inhibition of Lysine-Specific Demethylase-1 (LSD1/KDM1A) Promotes the Adipogenic Differentiation of hESCs Through H3K4 Methylation. Stem Cell Rev N/A:N/A (2016). WB . Read more (PubMed: 27059868) »

See all 7 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (rat choroid plexus cell line)
Specification
rat choroid plexus cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - Tritonx100 0.1% in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Mar 25 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line)
Specification
human glioblastoma cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X100 in PBS
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 11 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human glioblastoma cell lines)
Loading amount
100 µg
Specification
human glioblastoma cell lines
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Dec 23 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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