Anti-Histone H3 (phospho S10) antibody (ab47297)

Overview

  • Product name
    Anti-Histone H3 (phospho S10) antibody
    See all Histone H3 primary antibodies
  • Description
    Rabbit polyclonal to Histone H3 (phospho S10)
  • Specificity
    This antibody detects endogenous levels of Histone H3.1 only when phosphorylated at Serine 10.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IHC-Fr, WB, IHC-P, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Drosophila melanogaster
  • Immunogen

    Synthesized phosphopeptide derived from human Histone H3 around the phosphorylation site of serine 10 (R-K-SP-T-G)

  • Positive control
    • WB: Extracts from HeLa cells IHC-P: Human breast carcinoma tissue

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. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 59% PBS, 50% Glycerol, 0.87% Sodium chloride
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    The antibody was purified using epitope specific phosphopeptide. The antibody against non phosphopeptide was removed by chromatography using non phosphopeptide corresponding to the phosphorylation site.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab47297 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
Flow Cyt 1/500. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/1000.
IHC-Fr 1/2000. See Abreview.
WB 1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
IHC-P Use at an assay dependent concentration.
ELISA 1/10000.

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 (ab47297)

    Lane 1 : EGF + Calyculin treated HeLa cells + phospho-peptide.
    Lane 2 : EGF + Calyculin treated HeLa cells. No phosphopeptide.

    Lysates/proteins at 30 µg per lane.


    Predicted band size : 15 kDa
  • ab47297 detecting Histone H3 (phosphoS10) in assynchronous HeLa cells, in conjunction with a goat anti-rabbit secondary antibody conjugated to Cy3 (ab6939). Cells were also counterstained with DAPI in order to highlight the nucleus. For further details, please refer to abreview.

    See Abreview

  • ab47297 staining human breast carcinoma by IHC-P (left hand panel). The right hand panel shows staining in the presence of immunizing phosphopeptide.

  • ab47297 staining Histone H3 (phospho S10) in embryonic mouse brain by IHC-Fr (Paraformaldehyde-fixed Frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% TX-100 and blocked with 10% serum for 1 hour at 25°C. Samples were incubated with primary antibody 1/2000 (10%NGS in PBS + 0.1% TX10) for 16 hours at 25°C. A goat anti-rabbit Alexa Fluor®  488 (ab150077), dilution 1/400, was used as secondary antibody. The image shows ab47297 staining (green) in the CNS ventricular zone; there was no background staining with ab47297.

    See Abreview

  • ab47297 (Rabbit polyclonal to Histone H3 (phospho S10) ) identifying a sub-population of HCT116 cells with elevated FL1-H (PhosS10 labeling). For further experimental details please refer to abreview.

    See Abreview

References

This product has been referenced in:
  • Lim NR  et al. Glial-Specific Functions of Microcephaly Protein WDR62 and Interaction with the Mitotic Kinase AURKA Are Essential for Drosophila Brain Growth. Stem Cell Reports 9:32-41 (2017). IHC ; Drosophila melanogaster . Read more (PubMed: 28625535) »
  • Zhang B  et al. GSK3ß-Dzip1-Rab8 cascade regulates ciliogenesis after mitosis. PLoS Biol 13:e1002129 (2015). Read more (PubMed: 25860027) »

See all 11 Publications for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Human Cell lysate - whole cell (HCT116)
Specification
HCT116
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 19 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (human colon cancer cells)
Loading amount
25 µg
Specification
human colon cancer cells
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Apr 22 2013

Merci de nous avoir contactés.

Le kit ELISA ab100646 est en effet approprié pour le dosage du TGF-alpha humain contenu dans les supernageants de cultures cellulaires, urine, sérum et plasma.

Pour les anti-Histone ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (Human mammary cells)
Loading amount
50000 cells
Specification
Human mammary cells
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Ms. pierre laetitia

Verified customer

Submitted Feb 20 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Colon cell line)
Loading amount
20 µg
Specification
Colon cell line
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 29 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (embryonic mouse brain)
Specification
embryonic mouse brain
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TX-100 with Ab
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Dr. Ben Deverman

Verified customer

Submitted Jul 28 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (HCT116 (colorectal))
Specification
HCT116 (colorectal)
Fixation
Ethanol
Permeabilization
No
Gating Strategy
Cells were labeled with PI to reveal DNA content.
Username

Dr. Kirk Mcmanus

Verified customer

Submitted Oct 13 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

Submitted May 08 2007

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

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