Anti-Histone H3 (phospho S10) antibody (ab183626)

Overview

  • Product name
    Anti-Histone H3 (phospho S10) antibody
    See all Histone H3 primary antibodies
  • Description
    Rabbit polyclonal to Histone H3 (phospho S10)
  • Tested applications
    Suitable for: ChIP, IP, IHC - Wholemount, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Zebrafish
    Predicted to work with: Chimpanzee
  • Immunogen

    Synthetic peptide within Human Histone H3 (phospho S10). The exact sequence is proprietary. Synthetic phospho-peptide corresponding to amino acids residues surrounding the phospho Ser10.
    Database link: P68431

  • Positive control
    • U2OS (100ng/ml Nocodazole treatment for 24hr) , 4T1 (100ng/ml Nocodazole treatment for 24hr) , Rat2 whole cell lysate (100 ng/ml Nocodazole treatment for 24 hr); RFP-tag transfected HeLa cells; Mitotic HeLa cells; Human gastric carcinoma tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab183626 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 Use at an assay dependent concentration.
IP 1/100 - 1/500.
IHC - Wholemount 1/100 - 1/500.
IHC-P 1/100 - 1/1000.
WB 1/500 - 1/3000. Predicted molecular weight: 15 kDa.
ICC/IF 1/100 - 1/1000.

Target

  • Function
    Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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 throughout the cell cycle independently of DNA synthesis.
  • 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.
    Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. 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), which are linked to gene repression, are underrepresented. 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. Phosphorylation on Ser-32 (H3S31ph) is specific to regions bordering centromeres in metaphase chromosomes.
    Ubiquitinated. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H3 3 like sequence MH921 antibody
    • H3 3A antibody
    • H3 a antibody
    • H3 b antibody
    • H3 c antibody
    • H3 d antibody
    • H3 f antibody
    • H3 h antibody
    • H3 histone family member E pseudogene antibody
    • H3 i antibody
    • H3 j antibody
    • H3 k antibody
    • H3 l antibody
    • H33_HUMAN antibody
    • H3F3 antibody
    • H3f3b antibody
    • Histone H3 3 pseudogene antibody
    • Histone H3.3 antibody
    see all

Images

  • All lanes : Anti-Histone H3 (phospho S10) antibody (ab183626) at 1/1000 dilution

    Lane 1 : U2OS whole cell lysate (untreated)
    Lane 2 : U2OS whole cell lysate (serum deprivation for 48hr)
    Lane 3 : U2OS whole cell lysate (100ng/ml Nocodazole treatment for 24hr)

    Lysates/proteins at 30 µg per lane.


    Predicted band size : 15 kDa
  • Confocal immunofluorescent analysis of mitotic HeLa cells labeling Histone H3 (phospho S10) with ab183626 at 1/500 (green). Red: Tubulin, alpha 1b protein stained by TUBA1B antibody at 1/2500 . Blue: Hoechst 33342 staining.

  • Immunohistochemical analysis of paraffin embedded Human gastric carcinoma tissue labeling Histone H3 (phospho S10) with ab183626 at 1/500.

  • ab183626 immunoprecipitating Histone H3 in HeLa whole cell lysate.

    Lane (-): Untreated HeLa whole cell lysate (30 µg)

    Lane (+): Treated HeLa whole cell lysate (30 µg) were separated by 15% SDS-PAGE, and the membrane was blotted with ab183626 diluted at 1:2000.

    For western blotting,  HRP-conjugated anti-rabbit IgG was used as a secondary antibody. 

  • Cross-linked ChIP was performed with HeLa chromatin extract treated with Nocodazole (100 ng/ml for 24 h) and 5 μg of either control rabbit IgG or anti-Histone H3 (phospho S10) antibody (ab183626). The precipitated DNA was detected by PCR with primer set targeting to GAPDH promoter.

  • All lanes : Anti-Histone H3 (phospho S10) antibody (ab183626) at 1/500 dilution

    Lane 1 : Rat2 whole cell lysate
    Lane 2 : Rat2 whole cell lysate (100 ng/ml Nocodazole treatment for 24 hr)

    Lysates/proteins at 30 µg per lane.


    Predicted band size : 15 kDa
  • Immunofluorescent analysis of RFP-tag transfected HeLa cells labeling Histone H3 (phospho S10) with ab183626 at 1/500 (green). Red: Tubulin, alpha 1b protein stained by TUBA1B antibody at 1/500 . Blue: Hoechst 33342 staining.

  • All lanes : Anti-Histone H3 (phospho S10) antibody (ab183626) at 1/1000 dilution

    Lane 1 : 4T1 whole cell lysate
    Lane 2 : 4T1 whole cell lysate (100ng/ml Nocodazole treatment for 24hr)

    Lysates/proteins at 30 µg per lane.


    Predicted band size : 15 kDa
  • Immunohistochemistry (wholemount) analysis of zebrafish labelling Histone H3 (phopho S10) with ab183626 at a 1/100 dilution.

    Sample: Paraformaldehyde-fixed 2 day-post-fertilization zebrafish embryo.

  • Immunohistochemistry (wholemount) analysis of zebrafish labelling Histone H3 (phopho S10) with ab183626 at a 1/100 dilution.

    Sample: Paraformaldehyde-fixed 2 day-post-fertilization zebrafish embryo.

References

ab183626 has not yet been referenced specifically in any publications.

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