Anti-Histone H3 (tri methyl K9, phospho S10) antibody [mAbcam 14955] - BSA and Azide free (ab238673)

Overview

  • Product name

    Anti-Histone H3 (tri methyl K9, phospho S10) antibody [mAbcam 14955] - BSA and Azide free
  • Description

    Mouse monoclonal [mAbcam 14955] to Histone H3 (tri methyl K9, phospho S10) - BSA and Azide free
  • Host species

    Mouse
  • Specificity

    ab14955 recognises phospho S10 on Histone H3, even when K9 is tri methylated. It recognises a phospho S28 peptide by ELISA, but is not blocked by phospho S28 in WB.
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ELISAmore details
  • Species reactivity

    Reacts with: Mouse, Human, Xenopus laevis, Drosophila melanogaster, Indian muntjac, African green monkey
    Predicted to work with: Chicken, Saccharomyces cerevisiae, Caenorhabditis elegans, Schizosaccharomyces pombe, Zebrafish, Mammals, Tobacco, Chlamydomonas reinhardtii, Aspergillus nidulans, Neurospora crassa
  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (tri methyl K9, phospho S10) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.

  • Positive control

    • WB: Control HeLa Histone Prep. Colcemid treated HeLa Histone prep. HeLa cell lysate treated with calyculin A. IHC-P: Human kidney tissue.
  • General notes

    ab238673 is a PBS only version of ab14955.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising tri methyl K9 and phospho S10 dimodified peptide. Clones were also positively screened against both tri methyl K9 and phospho S10 peptides. Clones were negatively screened against the unmodified version of the peptides. This clone binds to the tri methyl K9 and phospho S10 dimodified peptide and to the phospho S10 peptide, but not to the tri methyl K9 peptide or to equivalent unmodified Histone H3 peptide.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    mAbcam 14955
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab238673 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 Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
IHC-P Use a concentration of 1 µg/ml.
Flow Cyt 1/1000.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ELISA Use at an assay dependent concentration.

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

    see all
  • 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 (tri methyl K9, phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955) at 1 µg/ml

    Lane 1 : Control HeLa Histone Prep
    Lane 2 : Colcemid treated HeLa Histone prep
    Lane 3 : Control HeLa Histone Prep with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
    Lane 4 : Colcemid treated HeLa Histone prep with Human Histone H3 (tri methyl K9, phospho S10) peptide (ab15644) at 1 µg/ml
    Lane 5 : Control HeLa Histone Prep with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
    Lane 6 : Colcemid treated HeLa Histone prep with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
    Lane 7 : Control HeLa Histone Prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 8 : Colcemid treated HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 9 : Control HeLa Histone Prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
    Lane 10 : Colcemid treated HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : Rabbit polyclonal to Mouse IgG H&L (HRP) at 1/3000 dilution

    Predicted band size: 15 kDa
    Observed band size: 17 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different formulation containing PBS and Azide (ab14955).

  • IHC image of ab14955 staining Histone H3 (phospho S10) in human kidney formalin fixed paraffin embedded tissue sections, performed on a Leica Bond.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab14955, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    This data was developed using the same antibody clone in a different formulation containing PBS and Azide (ab14955).

  • By ELISA, ab14955 detects: the singly modified phospho S10 peptide and the dual modified phospho S10 and tri methyl K9 peptide (the orange and grey lines respectively), less strongly detects the phospho S28 peptide (purple line), does not detect the equivalent non-modified Histone H3 peptide for S10 or the singly modified tri methyl K9 peptide (the 2 lines at the bottom of the figure).

    The antibody recognises phospho S28 by ELISA (although a phospho S28 peptide does not block the antibody in Western blotting).

    This data was developed using the same antibody clone in a different formulation containing PBS and Azide (ab14955).

  • All lanes : Anti-Histone H3 (tri methyl K9, phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955) at 0.5 µg/ml

    Lane 1 : Control HeLa Histone prep
    Lane 2 : Colecemid treated HeLa Histone prep
    Lane 3 : Control HeLa Histone prep with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg
    Lane 4 : Colecemid treated HeLa Histone prep with Human Histone H3 (unmodified ) peptide (ab7228) at 1 µg/ml
    Lane 5 : Control HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg/ml
    Lane 6 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S28) peptide (ab14793) at 1 µg
    Lane 7 : Control HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 8 : Colecemid treated HeLa Histone prep with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml

    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution

    Predicted band size: 15 kDa
    Observed band size: 20 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different formulation containing PBS and Azide (ab14955).

  • All lanes : Anti-Histone H3 (tri methyl K9, phospho S10) antibody [mAbcam 14955] - ChIP Grade (ab14955) at 1/2000 dilution

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : HeLa cell lysate treated with calyculin A
    Lane 3 : HeLa cell lysate treated with calyculin A and alkaline phosphatase

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/20000 dilution

    Predicted band size: 15 kDa



    This data was developed using the same antibody clone in a different formulation containing PBS and Azide (ab14955).

References

ab238673 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab238673.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up