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Epigenetics and Nuclear Signaling Histones H3 Phosphorylated
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Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)

  • Datasheet
  • SDS
Reviews (11)Q&A (1)References (74)

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Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
  • Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)

Key features and details

  • Rat monoclonal [HTA28] to Histone H3 (phospho S28)
  • Suitable for: WB, ICC
  • Reacts with: Human
  • Isotype: IgG2a

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Overview

  • Product name

    Anti-Histone H3 (phospho S28) antibody [HTA28]
    See all Histone H3 primary antibodies
  • Description

    Rat monoclonal [HTA28] to Histone H3 (phospho S28)
  • Host species

    Rat
  • Tested Applications & Species

    Application Species
    ICC
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3 aa 23-35 (phospho S28) conjugated to Keyhole Limpet Haemocyanin (KLH).
    Sequence:

    KAARKSA PATGGV


    Database link: P68431
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: HeLa (treated with Nocodazole) whole cell extract. ICC: HeLa cells.

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: 0.097% Sodium azide
    Constituents: 0.0268% PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified from culture supernatant of hybridoma cells grown in a bioreactor
  • Clonality

    Monoclonal
  • Clone number

    HTA28
  • Isotype

    IgG2a
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Phosphorylated

Associated products

  • Compatible Secondaries

    • Goat Anti-Rat IgG H&L (Alexa Fluor® 488) (ab150157)
    • Goat Anti-Rat IgG H&L (HRP) (ab205720)
  • Isotype control

    • Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (ab18450)
  • Recombinant Protein

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab10543 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
Human
WB
Human
All applications
Mouse
Hamster
Cow
Application Abreviews Notes
WB (1)
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 15 kDa.

We advise you to centrifuge this product vial before use.

ICC (2)
Use a concentration of 5 µg/ml.
Notes
WB
Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 15 kDa.

We advise you to centrifuge this product vial before use.

ICC
Use a concentration of 5 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
    • Omim: 602810 Human
    • SwissProt: P68432 Cow
    • SwissProt: P68431 Human
    • SwissProt: P68433 Mouse
    • 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
    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 S28) antibody [HTA28] (ab10543)
    Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
    Lane 1 : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 0.5 µg/ml
    Lane 2 : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 1 µg/ml
    Lane 3 : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 2 µg/ml

    All lanes : HeLa (treated with Nocodazole) whole cell extract

    Secondary
    All lanes : Goat Anti-Rat IgG-peroxidase conjugate

    Developed using the ECL technique.
  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
    Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)

    Immunocytochemical analysis of HeLa cells fixed and permeabilized with methanol followed by methanol:acetone. Histone H3 (phospho S28) was labeled with ab10543 at 5 μg/mL in ICC/IF (Green). The secondary antobody was a Goat Anti-Rat IgG-FITC conjugate. The nuclear counerstain was DAPI (Blue).

  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
    Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)This image is courtesy of an anonymous Abreview

    ab10543 staining Histone H3 (phospho S28) in Human neural progenitor cells from iPS cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 1% serum, 0.1% BSA in PBS for 30 minutes at room temperature. Samples were incubated with primary antibody (2ug/ml in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rat IgG2a polyclonal was used as the secondary antibody (1/500). Total cells were stained using DAPI (blue)

    See Abreview

  • Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
    Western blot - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
    All lanes : Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543) at 1 µg/ml

    Lane 1 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated
    Lane 2 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated with Human Histone H3 (unmodified ) peptide (ab2623) at 0.5 µg/ml
    Lane 3 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated with Human Histone H3 (phospho S28) peptide (ab5499) at 0.5 µg/ml
    Lane 4 : HeLa Histone Preparation Nuclear Lysate - Colcemid-treated with Human Histone H3 (phospho S10) peptide (ab11477) at 0.5 µg/ml

    Lysates/proteins at 2.5 µg per lane.

    Secondary
    All lanes : Peroxidase Conjugated AffiniPure Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab10543 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
    Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)This image is courtesy of an anonymous Abreview

    ab10543 staining Histone H3 (phospho S28) in Mouse neural stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 4% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS + 4% serum) for 16 hours at 4°C. An Alexa Fluor® 546-conjugated anti-rat polyclonal was used as the secondary antibody (1/500). DAPI is stainded blue

    See Abreview

  • Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)
    Immunocytochemistry - Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)This image is courtesy of an Abreview submitted by Dr Kirk McManus

    ab10543 staining HeLa cells by ICC/IF. The cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X100 in PBS. The cells were then stained with ab10543 at 1/1000 in PBS for 1h at 22°C. A goat anti-rat Alexa Fluro 488 (ab150157) at 1/200 was used as the secondary antibody. Nuclei are stained in red with DAPI. The antibody produces the expected mitotic-associated staining pattern and is extremely strong. MeOH fixed samples were also evaluated and produced a similar, strong staining pattern in mitotic cells.

Protocols

  • Flow cytometry 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 (74)

Publishing research using ab10543? Please let us know so that we can cite the reference in this datasheet.

ab10543 has been referenced in 74 publications.

  • Wei Y  et al. PDK1 regulates the survival of the developing cortical interneurons. Mol Brain 13:65 (2020). PubMed: 32366272
  • Zhang Y  et al. Contribution of Increased Expression of Yin Yang 2 to Development of Cardiomyopathy. Front Mol Biosci 7:35 (2020). PubMed: 32195266
  • Monroe TO  et al. YAP Partially Reprograms Chromatin Accessibility to Directly Induce Adult Cardiogenesis In Vivo. Dev Cell 48:765-779.e7 (2019). PubMed: 30773489
  • Bonnefont J  et al. Cortical Neurogenesis Requires Bcl6-Mediated Transcriptional Repression of Multiple Self-Renewal-Promoting Extrinsic Pathways. Neuron 103:1096-1108.e4 (2019). PubMed: 31353074
  • Saito K  et al. Dorsal-to-Ventral Cortical Expansion Is Physically Primed by Ventral Streaming of Early Embryonic Preplate Neurons. Cell Rep 29:1555-1567.e5 (2019). PubMed: 31693895
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 12 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-Histone H3 (phospho S28) antibody [HTA28]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Petaurus breviceps Tissue sections (Skin tissue)
Permeabilization
Yes - Triton
Specification
Skin tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Oct 12 2017

IHC - Wholemount abreview for Anti-Histone H3 (phospho S28) antibody [HTA28]

Excellent
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Drosophila melanogaster Embryo (wholemount)
Specification
wholemount
Read More

Abcam user community

Verified customer

Submitted Jan 10 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (phospho S28) antibody [HTA28]

Average
Abreviews
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Sample
Human Cell (human)
Specification
human
Permeabilization
No
Fixative
Parafilm
Read More

Ms. Nikita Wright

Verified customer

Submitted Sep 29 2014

Immunocytochemistry abreview for Anti-Histone H3 (phospho S28) [HTA28] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Mar 27 2014

Western blot abreview for Anti-Histone H3 (phospho S28) [HTA28] antibody

Average
Abreviews
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 Mar 04 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (phospho S28) [HTA28] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
1% serum, 0.1% triton, 0.1% BSA in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Sample
Human Cell (human neural progenitor cells from ips cell line)
Specification
human neural progenitor cells from ips cell line
Permeabilization
Yes - see blocking buffer
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jan 27 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (phospho S28) [HTA28] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde
Read More

Dr. Kirk Mcmanus

Verified customer

Submitted Oct 03 2013

Question

Im interested in follow H3S28phosphorilation by ChIP in Saccharomyces cerevisiae. Do you have any antibody?

Read More

Abcam community

Verified customer

Asked on Feb 13 2013

Answer

ab10543 shares between 76-92% homology with the yeast protein so both could work in yeast but this has not been tested and therefore we cannot guarantee this. If you wish to test one of this antibody, I could give you a testing discount using our AbTrial program.

Read More

Abcam Scientific Support

Answered on Feb 13 2013

Immunohistochemistry (Frozen sections) abreview for Anti-Histone H3 (phospho S28) antibody [HTA28]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Neonatal Heart)
Specification
Neonatal Heart
Fixative
Paraformaldehyde
Permeabilization
Yes - Saponin 0,1%
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Read More

Abcam user community

Verified customer

Submitted May 11 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (phospho S28) antibody [HTA28]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse embryonic fibroblast (MEF))
Specification
mouse embryonic fibroblast (MEF)
Fixative
Paraformaldehyde
Permeabilization
Yes - -20C ethanol
Read More

Abcam user community

Verified customer

Submitted Aug 19 2009

1-10 of 12 Abreviews or Q&A

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