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Product name
Anti-Histone H3 (phospho S10) antibody - Mitosis Marker
See all Histone H3 products (20) ...
Description
Rabbit polyclonal to Histone H3 (phospho S10) - Mitosis Marker
Specificity
This antibody is specific for phosho S10 of histone H3. We believe that it does not recognise the non-modified histone - no blocking is seen with the non-phospho peptide (see Western blot image).
Tested applications
ICC/IF, WB, IHC-P, IHC - Wholemount, IHC-Fr, Flow Cyt, ChIP, IHC-FoFrmore details
Cross reactivity
Reacts with
Mouse, Cow, Human, Fruit fly (Drosophila melanogaster), Indian Muntjac, Zebrafish, African Green Monkey
Predicted to work with
Rat, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Schizosaccharomyces pombe, Neurospora crassa
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, phosphorylated at S10.
(Peptide available as ab114 77.)
Positive control
Colcemid-treated HeLa histone Preparation; Hela whole cell extract
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Phosphorylated
Applications
Show applications key
Our Abpromise guarantee covers the use of ab5176 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: Use at an assay dependent dilution.
WB: 1/500 - 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).Can be blocked with Histone H3 peptide - phospho S10 (ab11477).
IHC-P: Use at an assay dependent dilution. IHC-Wholemount. IHC-Methanol: Use at an assay dependent concentration (PMID 20144789).
IHC - Wmt: Use at an assay dependent concentration.
IHC-Fr: 1/200. (see Abreview)
Flow Cyt: 1/1000.
ChIP: Use at an assay dependent dilution. PubMed: 20089855
IHC-FoFr: Use at an assay dependent dilution. PubMed: 19622634
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).
Alternative names
- H3 3 like sequence MH921 antibody
- H3 3 like sequence MH921 antibody
- H3 3A antibody
see all
Database links
- Entrez Gene: 176359 Caenorhabditis elegans
- Entrez Gene: 326601 Cow
- Entrez Gene: 45314 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 31848 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 33736 Fruit fly (Drosophila melanogaster)
- Entrez Gene: 3020 Human
- Entrez Gene: 3021 Human
- Entrez Gene: 15078 Mouse
see all
Anti-Histone H3 (phospho S10) antibody - Mitosis Marker images:
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (phospho S10) antibody - Mitosis Marker (ab5176)
This image was kindly supplied by Prof Bryan Turner, University of Birmingham. Female Human Lymphoblastoid cells were incubated with ab5176.
Image courtesy of Dr. Bryan Turner, United Kingdom
Western blot - Histone H3 (phospho S10) antibody (ab5176)
All lanes : Anti-Histone H3 (phospho S10) antibody - Mitosis Marker (ab5176) at 1 µg/ml
Lane 1 : untreated histones
Lane 2 : colcemid treated histones
Lane 3 : untreated histones with
Lane 4 : colcemid treated histones with
Lane 5 : untreated histones with
Lane 6 : colcemid treated histones with
Lysates/proteins at 0.5 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP) (ab6721) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 17 kDa
Observed band size : 15-20 kDa (why is the actual band size different from the predicted?)
Exposure time : 10 seconds
Immunofluorescence - Histone H3 (phospho S10) antibody - Mitosis Marker (ab5176)
SKN cells stained with ab5176 (green) at a dilution of 1/100. The cells were fixed in paraformaldehyde for 10 minutes prior to incubation with ab5176. The DNA is stained with DAPI (blue). 100x magnification.
Darin McDonald, Hendzel Laboratory
Immunocytochemistry/ Immunofluorescence - Histone H3 (phospho S10) antibody - Mitosis Marker (ab5176)
ab5176 at a 1/100 dilution staining Drosophila melanogaster (wild type) polytene chromosomes by ICC/IF. The cells were formaldehyde fixed and blocked with 1% BSA prior to incubation with the antibody for 12 hours. Bound antibody was detected using a Cy3 conjugated goat anti-rabbit antibody.
This image is courtesy of an Abreview submitted by Miss Anita Ciurciu
Flow Cytometry - Histone H3 (phospho S10) antibody - Mitosis Marker (ab5176)
ab5176 (1/1000) staining a population of human HeLa cells positive for Histone H3 (phospho S10). Cells were trypsizined, pelleted and fixed in ice cold Ethanol. Debris was eliminated and FL2-A/FL2-W was used to eliminate clumping cells. For further experimental details please refer to abreview folder.
This image is courtesy of an Abreview submitted by Dr Kirk McManus
IHC - Wholemount - Anti-Histone H3 (phospho S10) antibody - Mitosis Marker (ab5176)
IHC-Wholemount image of Histone H3 (phospho S10) ab5176 on zebrafish jaw. 5 day old zebrafish were permeabilized with proteinase K (10ug/ml, 50min, 37degrees), prior to staining. Staining visible in mitotic cells. Image shows 2 cells labelled with H3 phospho S10 antibody (green), nuclei are labelled with DAPI (blue).
This image was taken from an abreview by Chrissy Hammond.
References for Anti-Histone H3 (phospho S10) antibody - Mitosis Marker (ab5176)
This product has been referenced in:
- Kiyomitsu T et al. Protein interaction domain mapping of human kinetochore protein Blinkin reveals a consensus motif for binding of spindle assembly checkpoint proteins Bub1 and BubR1. Mol Cell Biol 31:998-1011 (2011).Read more (PubMed: 21199919) »
- Kannan RP et al. Hepatitis C virus infection causes cell cycle arrest at the level of initiation of mitosis. J Virol 85:7989-8001 (2011). WB; Human.Read more (PubMed: 21680513) »
See all 21 publications for this product
Publishing research using ab5176? 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"