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Epigenetics and Nuclear Signaling Histones H3 Phosphorylated
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Anti-Histone H3 (phospho T11) antibody (ab5168)

  • Datasheet
  • SDS
Reviews (3)Q&A (2)References (17)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho T11) antibody (ab5168)
  • Western blot - Anti-Histone H3 (phospho T11) antibody (ab5168)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho T11) antibody (ab5168)
  • Western blot - Anti-Histone H3 (phospho T11) antibody (ab5168)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho T11) antibody (ab5168)

Key features and details

  • Rabbit polyclonal to Histone H3 (phospho T11)
  • Suitable for: WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Cow, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Histone H3 (phospho T11) antibody
    See all Histone H3 primary antibodies
  • Description

    Rabbit polyclonal to Histone H3 (phospho T11)
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Cow
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3 aa 1-100 (phospho T11) conjugated to keyhole limpet haemocyanin.
    Database link: P68431
    (Peptide available as ab24444)

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 9.97% L-Arginine

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Phosphorylated

Associated products

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Control Peptide

    • Human Histone H3 (phospho S10) peptide (ab11477)
    • Human Histone H3 (unmodified) peptide (ab7228)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human Histone H3 protein (Active) (ab198757)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab5168 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Cow
Human
All applications
Rat
Saccharomyces cerevisiae
Xenopus laevis
Arabidopsis thaliana
Caenorhabditis elegans
Drosophila melanogaster
Schizosaccharomyces pombe
Zebrafish
Neurospora crassa
Application Abreviews Notes
WB (3)
1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 µg/ml.
Notes
WB
1/500 - 1/1000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 µg/ml.

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.
  • Target information above from: UniProt accession P68431 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8350 Human
    • Entrez Gene: 8351 Human
    • Entrez Gene: 8352 Human
    • Entrez Gene: 8353 Human
    • Entrez Gene: 8354 Human
    • Entrez Gene: 8355 Human
    • Entrez Gene: 8356 Human
    • Entrez Gene: 8357 Human
    • Entrez Gene: 8358 Human
    • Entrez Gene: 8968 Human
    • Entrez Gene: 319152 Mouse
    • Entrez Gene: 319153 Mouse
    • Entrez Gene: 360198 Mouse
    • Entrez Gene: 97908 Mouse
    • Entrez Gene: 100364501 Rat
    • Entrez Gene: 100365669 Rat
    • Entrez Gene: 291159 Rat
    • Entrez Gene: 679994 Rat
    • Entrez Gene: 680511 Rat
    • Entrez Gene: 682330 Rat
    • Omim: 602810 Human
    • SwissProt: P68432 Cow
    • SwissProt: P68431 Human
    • SwissProt: P68433 Mouse
    • SwissProt: Q6LED0 Rat
    • Unigene: 132854 Human
    • Unigene: 247813 Human
    • Unigene: 247814 Human
    • Unigene: 248176 Human
    • Unigene: 443021 Human
    • Unigene: 484990 Human
    • Unigene: 532144 Human
    • Unigene: 533292 Human
    • Unigene: 546315 Human
    • Unigene: 586261 Human
    • Unigene: 591778 Human
    • Unigene: 221301 Mouse
    • Unigene: 261657 Mouse
    • Unigene: 377874 Mouse
    • Unigene: 390558 Mouse
    • Unigene: 397328 Mouse
    • Unigene: 138090 Rat
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho T11) antibody (ab5168)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (phospho T11) antibody (ab5168)
    IHC image of Histone H3 (phospho T11) staining in human normal testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab5168, 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 haematoxylin and mounted with DPX.

    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.
  • Western blot - Anti-Histone H3 (phospho T11) antibody (ab5168)
    Western blot - Anti-Histone H3 (phospho T11) antibody (ab5168)
    Anti-Histone H3 (phospho T11) antibody (ab5168) at 1/500 dilution + Calf thymus histone lysate

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution

    Performed under reducing conditions.

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



    Rabbit polyclonal to Histone H3 (phospho T11) - ab5168 at 1/500 dilution.

    Lane 1 contains 1ug calf thymus histone prep.

    Secondary ab: Goat anti-rabbit IgG HRP conjugate ab6721 (1/2000)

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho T11) antibody (ab5168)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho T11) antibody (ab5168)This image is courtesy of Darin McDonald

    SKN-SH cells were fixed in 4% paraformaldehyde for 10 mins, permeabilized in PBS-0.5% Triton X-100 for 5 mins and incubated for 30 minutes with ab5168 (1/100). The slides were rinsed once in PBS-Triton (0.1%), twice in PBS then incubated with the secondary antibody for 30 mins. The DNA is stained with DAPI (blue). Clear nuclear staining with ab5168 can be seen (green). 100x magnification.

  • Western blot - Anti-Histone H3 (phospho T11) antibody (ab5168)
    Western blot - Anti-Histone H3 (phospho T11) antibody (ab5168)
    All lanes : Anti-Histone H3 (phospho T11) antibody (ab5168) at 1 µg/ml

    Lane 1 : Colcemid treated HeLa Histone prep at 5 µg
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab27251) at 20 µg
    Lane 3 : Colcemid treated HeLa Histone prep at 5 µg with Human Histone H3 (phospho T11) peptide (ab24444) at 1 µg/ml
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab27251) at 20 µg with Human Histone H3 (phospho T11) peptide (ab24444) at 1 µg/ml
    Lane 5 : Colcemid treated HeLa Histone prep at 5 µg with Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/ml
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab27251) at 20 µg with Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/ml
    Lane 7 : Colcemid treated HeLa Histone prep at 5 µg with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml
    Lane 8 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate (ab27251) at 20 µg with Human Histone H3 (phospho S10) peptide (ab11477) at 1 µg/ml

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution

    Performed under reducing conditions.

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



    This antibody is blocked by the Histone H3 phospho T11 peptied but not by the respective unmodified peptide or the Histone H3 phospho S10 peptide. This confirms that the antibody is specific for phospho T11 of Histone H3.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho T11) antibody (ab5168)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho T11) antibody (ab5168)
    ICC/IF image of ab5168 stained Hek293 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab5168, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa and HepG2 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (17)

Publishing research using ab5168? Please let us know so that we can cite the reference in this datasheet.

ab5168 has been referenced in 17 publications.

  • Karakkat JV  et al. The metabolic sensor PASK is a histone 3 kinase that also regulates H3K4 methylation by associating with H3K4 MLL2 methyltransferase complex. Nucleic Acids Res 47:10086-10103 (2019). PubMed: 31529049
  • Oh S  et al. Histone H3 threonine 11 phosphorylation by Sch9 and CK2 regulates chronological lifespan by controlling the nutritional stress response. Elife 7:N/A (2018). PubMed: 29938647
  • Ginno PA  et al. Cell cycle-resolved chromatin proteomics reveals the extent of mitotic preservation of the genomic regulatory landscape. Nat Commun 9:4048 (2018). PubMed: 30279501
  • Zhang F & Guo F Effect of transplantation of cardiac stem cells overexpressing integrin-linked kinase on cardiac function of rats with acute myocardial infarction. Exp Ther Med 16:746-750 (2018). PubMed: 30116329
  • Yang CS  et al. The protein kinase C super-family member PKN is regulated by mTOR and influences differentiation during prostate cancer progression. Prostate 77:1452-1467 (2017). PubMed: 28875501
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Western blot abreview for Anti-Histone H3 (phospho T11) antibody

Good
Abreviews
Abreviews
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse ES Cells)
Gel Running Conditions
Reduced Denaturing (15% SDS PAGE)
Loading amount
50 µg
Specification
Mouse ES Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Mr. Utkarsh Kapoor

Verified customer

Submitted Dec 26 2012

Western blot abreview for Anti-Histone H3 (phospho T11) antibody

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (fibroblast)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
22 µg
Specification
fibroblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Aug 03 2010

Western blot abreview for Anti-Histone H3 (phospho T11) antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HT29 tumour cells)
Gel Running Conditions
Reduced Denaturing (16% Tris Glycine)
Loading amount
50 µg
Treatment
50nM SN38 for 6h +/- 50nM staurosporine
Specification
HT29 tumour cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted May 28 2009

Question

I received an inquiry from a customer who purchase ab5168 (Histone H3 (phospho T11)). The customer plans to perform WB for cell extract of mouse and yeast with the Ab, but she does not have enough experience. Do you have any advises/information/inputs about such experiment?

Read More

Abcam community

Verified customer

Asked on Feb 27 2007

Answer

I would recommend that your customer performs a histone extraction prior to performing western blotting. This would serve as an excellent positive control for the purpose of detection. I would direct them to the following protocol for the extraction of histones from cultured mouse cells; https://www.abcam.com/ps/pdf/protocols/Histone%20extraction%20protocol.pdf I would then direct them to our lab protocol for the testing of histones, available at the following link; https://www.abcam.com/ps/pdf/protocols/Histone%20western%20blotting.pdf This is an excellent place to start for the application of our antibodies against modified histones.

Read More

Abcam Scientific Support

Answered on Feb 27 2007

Question

I have a cusomter who is looking for a suitable positive control for ab5168. Unfortunately C2C12 cell line will not be suitable- please refer to her reply: I was hoping for the MuSK transfected COS cells from abcam as the C2C12 cell line is what I am working on and I'm looking for changes in MuSK in response to treatment. I don't want to use this line as my positive control since it's my experimental cell line. Would you have any suggestions?

Read More

Abcam community

Verified customer

Asked on May 11 2006

Answer

Whilst we do not have a recommended positive control for this antibody. I would like to recommend that a calf thymus histone lysate is used. This is shown in the western blot on the datasheet. Alternatively histones can be extracted and stored as aliquots at -20oC. For an easy histone extraction protocol please see below: https://www.abcam.com/assets/pdf/protocols/Histone%20extraction%20protocol.pdf

Read More

Abcam Scientific Support

Answered on May 15 2006

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