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Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)

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Overview

Product name

Anti-Histone H3 (phospho S28) antibody [HTA28]
See all Histone H3 products (20) ...

Description

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

Specificity

Does not detect the unphosphorylated epitope. It detects the phosphorylated histone molecule at the onset of mitosis (prophase, metaphase and weaker at the beginning of anaphase), but not during late anaphase.

Tested applications

IHC-Fr, ICC/IF, WB, ICC, Flow Cytmore details

Cross reactivity

Reacts with

Mouse, Chicken, Hamster, Cow, Human, Oikopleura - a pelagic tunicate, Marmoset (common)

Immunogen

Synthetic peptide conjugated to KLH, corresponding to amino acids 23-35 of Human Histone H3.

Positive control

Whole extract of cultured human acute T cell leukemia Jurkat cells, treated with Nocodazole.

Properties

Form

Liquid

Storage instructions

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Storage buffer

Preservative: 15mM Sodium Azide
Constituents: 1% BSA, 0.01M PBS, pH 7.4

Concentration

Concentration information loading...

Purity

Immunogen affinity purified

Clonality

Monoclonal

Clone number

HTA28

Isotype

IgG2a

Research areas

Epigenetics and Nuclear Signaling >> Histones >> H3 >> Phosphorylated

  • Immunohistochemistry (Frozen sections) - Histone H3 (phospho S28) antibody [HTA28] (ab10543)Immunohistochemistry (Frozen sections) - Histone H3 (phospho S28) antibody [HTA28] (ab10543) image (enlarge)

  • Immunocytochemistry/ Immunofluorescence - Histone H3 (phospho S28) antibody [HTA28] (ab10543)Immunocytochemistry/ Immunofluorescence - Histone H3 (phospho S28) antibody [HTA28] (ab10543) image (enlarge)

Applications

Show applications key

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.

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.

Target information above from: UniProt accessionP84243 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

Alternative names

  • H3 3 like sequence MH921 antibody
  • H3 3 like sequence MH921 antibody
  • H3 3A 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 histone family member E pseudogene antibody
  • H3 i antibody
  • H3 j antibody
  • H3 k antibody
  • H3 l antibody
  • H33_HUMAN antibody
  • H3F3 antibody
  • H3F3 antibody
  • H3f3b antibody
  • Histone H3 3 pseudogene antibody
  • Histone H3 3 pseudogene antibody
  • Histone H3.3 antibody
see all

Database links

see all

Anti-Histone H3 (phospho S28) antibody [HTA28] images:

  Immunohistochemistry (Frozen sections) - Histone H3 (phospho S28) antibody [HTA28] (ab10543)

Immunohistochemistry (Frozen sections) - Histone H3 (phospho S28) antibody [HTA28] (ab10543)

ab10543 staining Histone H3 (phospho S28) in mouse embryonic brain tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with paraformaldehyde and blocking with 10% serum for 1 hour at 250C was performed. The sample was incubated with primary antibody (1/300)  at 40C for 16 hours. An Alexa Fluor®488-conjugated Goat polyclonal to rat IgG was used as secondary antibody at 1/500 dilution.

This image is a courtesy of Anonymous Abreview

See Abreview

  Immunocytochemistry/ Immunofluorescence - Histone H3 (phospho S28) antibody [HTA28] (ab10543)

Immunocytochemistry/ Immunofluorescence - Histone H3 (phospho S28) antibody [HTA28] (ab10543)

ab10543 staining Histone H3 (phospho S28) in murine embryonic fibroblast cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in paraformaldehyde and permeabilised in -20°C ethanol. Samples were then incubated with primary antibody at a 1/200 dilution for 1 hour in 0.1%Saponin/ 2.5% BSA/ PBS. The secondary antibody used was a goat anti-rat IgG (H+L) conjugated to DyLight® 649 (red) used at a 1/300 dilution. Gamma tubulin (blue), Ki67 (green) using ab15580.

Image courtesy of an anonymous Abreview.

References for Anti-Histone H3 (phospho S28) antibody [HTA28] (ab10543)

This product has been referenced in:

  • Campsteijn C  et al. Expansion of cyclin D and CDK1 paralogs in Oikopleura dioica, a chordate employing diverse cell cycle variants. Mol Biol Evol 29:487-502 (2012). Oikopleura - a pelagic tunicate.Read more (PubMed: 21734012) »
  • Kelava I  et al. Abundant Occurrence of Basal Radial Glia in the Subventricular Zone of Embryonic Neocortex of a Lissencephalic Primate, the Common Marmoset Callithrix jacchus. Cereb Cortex : (2011). IHC-Fr; Marmoset (common).Read more (PubMed: 22114084) »

See all 4 publications for this product

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

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