RecombinantRabMAb

Recombinant Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

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Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • Immunoprecipitation - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • Dot Blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
  • Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E173] to Histone H3 (phospho S10 + T11)
  • Suitable for: ELISA, Dot blot, ICC/IF, WB, IHC-P, IP
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free HRP

Overview

  • Product name

    Anti-Histone H3 (phospho S10 + T11) antibody [E173]
    See all Histone H3 primary antibodies

  • Description

    Rabbit monoclonal [E173] to Histone H3 (phospho S10 + T11)

  • Host species

    Rabbit

  • Specificity

    This antibody detects Histone H3 phosphorylated on both Serine 10 and Threonine 11. However, the antibody shows higher affinity for phosphorylated Serine 10 than for phosphorylated Threonine 11. This was validated by ELISA, Dot Blot and WB peptide blocking experiments.

  • Tested applications

    Suitable for: ELISA, Dot blot, ICC/IF, WB, IHC-P, IPmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Drosophila melanogaster, a wide range of other species

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa whole cell lysate (ab150035) treated with Calyculin A. IHC-P: Lymphoma tissue. ICC/IF: HeLa cells IP: HeLa treated with calyculin A whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    E173

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32107 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
ELISA
Use at an assay dependent concentration.
Dot blot
1/1000.
ICC/IF (1)
1/250 - 1/1000.
WB (1)
1/2500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
IHC-P (2)
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/50.
Notes
ELISA
Use at an assay dependent concentration.
Dot blot
1/1000.
ICC/IF
1/250 - 1/1000.
WB
1/2500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/50.
Application notes
Is unsuitable for Flow Cyt.

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

    see all

  • 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

  • Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/10000 dilution

    Lane 1 : Untreated HeLa 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 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


    Exposure time: 1 second


    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000, 2 μg/mL) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] (1/200, 2.5 μg/mL) - Microtubule Marker (Alexa Fluor® 594). DAPI (blue) was used as a nuclear counterstain.

    Confocal image showing nuclear staining on mitotic HeLa cell.

  • Immunoprecipitation - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Immunoprecipitation - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

    Purified ab32107 at 1/50 dilution (2µg) immunoprecipitating Histone H3 in HeLa treated with calyculin A whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) treated with calyculin A whole cell lysate 10µg
    Lane 2 (+): ab32107 + HeLa treated with calyculin A whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32107 in HeLa treated with calyculin A whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 17 kDa

  • Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Western blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    All lanes : Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107) at 1/1000 dilution

    Lane 1 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho S10 peptide
    Lane 2 : HeLa whole cell lysate -treated with Calyculin A with Histone H3 phospho T11 peptide
    Lane 3 : HeLa whole cell lysate -treated with Calyculin A with unmodified Histone H3 peptide
    Lane 4 : HeLa whole cell lysate -treated with Calyculin A with no peptide

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 1 second


    Blocking/Diluting buffer 5% NFDM/TBST

  • Dot Blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Dot Blot - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

    Dot blot analysis of Histone H3 phospho S10 peptide (Lane 1), Histone H3 phospho T11 peptide (Lane 2) and Histone H3 unmodified peptide (Lane 3) labelling Histone H3 (phospho S10 + T11) with ab32107 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) was used as the secondary antibody at a dilution of 1/10 000.

    Blocking and diluting buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

    Ab32107, at a 1/50 dilution, staining Histone H3 (phospho S10 + T11) in paraffin embedded lymphoma tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)This image is courtesy of an Abreview submitted by Dr Kirk McManus

    ab32107 (1/1000) staining Histone H3 (phospho S10 + T11) in paraformaldehyde-fixed, DAPI counterstained HeLa cells. Secondary antibody: Goat anti-Rabbit IgG conjugated to Cy3® (1/200). Please refer to abreview for further details.

    See Abreview

  • ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

    Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

    Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (100 ng/mL).

    Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).

  • ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    ELISA - Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

    Direct ELISA antibody dose-response curve using ab32107 (0-1000 ng/mL).

    Peptides - Histone H3 unmodified peptide, Histone H3 phospho S10 peptide, Histone H3 phospho T11 peptide (10 ng/mL).

    Secondary antibody - Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1/2500).

  • Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)
    Anti-Histone H3 (phospho S10 + T11) antibody [E173] (ab32107)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (41)

Publishing research using ab32107? Please let us know so that we can cite the reference in this datasheet.

ab32107 has been referenced in 41 publications.

  • Wu Y  et al. LRP6 downregulation promotes cardiomyocyte proliferation and heart regeneration. Cell Res 31:450-462 (2021). PubMed: 32973339
  • Niu K  et al. Canonical and early lineage-specific stem cell types identified in planarian SirNeoblasts. Cell Regen 10:15 (2021). PubMed: 33740162
  • Bohr TE  et al. Planarian stem cells sense the identity of the missing pharynx to launch its targeted regeneration. Elife 10:N/A (2021). PubMed: 34156924
  • Shiroor DA  et al. Injury Delays Stem Cell Apoptosis after Radiation in Planarians. Curr Biol 30:2166-2174.e3 (2020). PubMed: 32386527
  • Lin TY  et al. BIRC5/Survivin is a novel ATG12-ATG5 conjugate interactor and an autophagy-induced DNA damage suppressor in human cancer and mouse embryonic fibroblast cells. Autophagy 16:1296-1313 (2020). PubMed: 31612776
View all Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (phospho S10 + T11) antibody [E173]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cat Tissue sections (Skin tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: BH6 citrate buffer
Permeabilization
No
Specification
Skin tumor
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 24°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Nov 24 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Histone H3 (phospho S10 + T11) antibody [E173]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Skin, tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Specification
Skin, tumor
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 20°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Oct 01 2020

Western blot abreview for Anti-Histone H3 (phospho S10) antibody [E173]

 Excellent
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - whole (0-4hr embryos)
Specification
0-4hr embryos
Treatment
control shRNA and wee shRNA embryos
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Read More

DR. Richelle Sopko

Verified customer

Submitted Dec 04 2013

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (phospho S10) antibody [E173]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Read More

DR. Kirk Mcmanus

Verified customer

Submitted Feb 20 2007

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