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Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

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Overview

Product name

Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade
See all Histone H3 products (17) ...

Description

Mouse monoclonal [mAbcam 10799] to Histone H3 - ChIP Grade

Tested applications

ChIP, WB, IHC-Pmore details

Cross reactivity

Reacts with

Mouse, Human, Fruit fly (Drosophila melanogaster), Monkey

Predicted to work with

a wide range of other species, all Mammals

Immunogen

Synthetic peptide derived from within residues 100 to the C-terminus of Human Histone H3. Generated with the same immunogen as our successful rabbit polyclonal ab1791.

Read Abcam's proprietary immunogen policy

Positive control

This antibody gave a positive result in the following lysates: HeLa Whole Cell, Calf Thymus Histone and NIH3T3 (Mouse Embryonic Fibroblast) Whole Cell.

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: 0.02% Sodium Azide
Constituents: PBS, pH 7.5

Concentration

Concentration information loading...

Purity

IgG fraction

Clonality

Monoclonal

Clone number

mAbcam 10799

Myeloma

Sp2/0-Ag14

Isotype

IgG3

Light chain type

kappa

Research areas

Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Unmodified

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) image (enlarge)

  • ChIP - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)ChIP - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) image (enlarge)

  • Western blot - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)Western blot - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) image (enlarge)

Applications

Show applications key

Our Abpromise guarantee covers the use of ab10799 in the following tested applications.

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

Application notes

ChIP: Use 2-4µg for 25µg of chromatin
IHC-P: Use at a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 18 kDa (predicted molecular weight: 15 kDa). Can be blocked with Histone H3 peptide (ab12149).

Is unsuitable for ICC/IF.

Not yet tested in other applications.
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 histone antibody
  • H3 histone family member E pseudogene antibody
  • H3 histone family member E pseudogene antibody
  • H33_HUMAN antibody
  • H3F3 antibody
  • H3f3b antibody
  • HIST3H3 antibody
  • Histone H3 3 pseudogene antibody
  • Histone H3 3 pseudogene antibody
  • Histone H3.3 antibody
see all

Database links

see all

Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade images:

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

IHC image of Histone H3 staining in human breast carcinoma FFPE section, performed on a 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 ab10799, 5µg/ml, for 8 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.

  ChIP - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

ChIP - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 2µg of ab10799 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.    

  Western blot - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

Western blot - Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

All lanes : Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799) at 1 µg/ml

Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 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?)


Exposure time : 90 seconds

References for Anti-Histone H3 antibody [mAbcam 10799] - ChIP Grade (ab10799)

This product has been referenced in:

  • Marban Cet al. Genome-Wide Binding Map of the HIV-1 Tat Protein to the Human Genome. PLoS One 6:e26894 (2011). WB; Human.Read more (PubMed: 22073215) »
  • Peng Het al. Transcriptional coactivator HCF-1 couples the histone chaperone Asf1b to HSV-1 DNA replication components. Proc Natl Acad Sci U S A 107:2461-6 (2010). WB; Monkey.Read more (PubMed: 20133788) »

See all 7 publications for this product

Publishing research using ab10799? 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"