Overview

  • Product name

    Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade
    See all Histone H3 primary antibodies
  • Description

    Rabbit polyclonal to Histone H3 (mono methyl K4) - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ChIP, ICC/IF, PepArr, Dot blot, CHIPseqmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, a wide range of other species
  • Immunogen

    Synthetic peptide within Human Histone H3 (mono methyl K4) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary. Peptide surrounding K4.

  • Positive control

    • WB: HeLa whole cell extract and Histone extract. ICC/IF: HeLa cells. ChIP and ChIP seq: chromatin from K562 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab195391 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
WB 1/500.
ChIP Use 1-10µg for 105 cells.
ICC/IF 1/200.
PepArr 1/2000.
Dot blot 1/5000.
CHIPseq Use 1µg for 105 cells.

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 assays were performed using human K562 cells, ab195391 and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration consisting of 1, 2, 5 and 10 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for a region surrounding the ACTB and GAS2L1 gene, respectively, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. Image shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

  • ChIP was performed on sheared chromatin from 100,000 K562 cells using 1 µg of ab195391. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Image A and B shows the H3K4me1 signal in two genomic regions containing the ACTB and GAS2L1 positive controls. The position of the amplicon used for ChIP-qPCR is indicated by an arrow. Image C shows the H3K4me1 peak distribution along a 1 Mb genomic region of chromosome 5.

  • To test the cross reactivity of the ab195391, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified Histone H3 K4 (H3K4). One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1/5000. Image shows a high specificity of the antibody for the modification of interest.

  • All lanes : Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab195391) at 1/500 dilution

    Lane 1 : HeLa whole cell extract at 25 µg
    Lane 2 : Histone extracts from HeLa cells at 15 µg
    Lane 3 : Recombinant histone H2A at 1 µg
    Lane 4 : Recombinant histone H2B at 1 µg
    Lane 5 : Recombinant histone H3 at 1 µg
    Lane 6 : Recombinant histone H4 at 1 µg


    TBS-Tween containing 5% skimmed milk.

  • Immunocytochemistry/ Immunofluorescence of HeLa cells labeling Histone H3 (mono methyl K4) using ab195391 at a 1/200 dilution.

    Cells were fixed with 4% formaldehyde for 10 minutes and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the ab195391 (left) diluted 1/200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The right panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown below.

References

ab195391 has not yet been referenced specifically in any publications.

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