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Products:Epigenetics and Nuclear Signaling >> Histones >> H3 >> Unmodified
Overview
Product name
Anti-Histone H3 antibody - ChIP Grade
See all Histone H3 products (17) ...
Description
Goat polyclonal to Histone H3 - ChIP Grade
Tested applications
ChIP, IP, ChIP/Chip, WBmore details
Cross reactivity
Reacts with
Mouse, Chicken, Dog, Human, Saccharomyces cerevisiae, Xenopus laevis, Fruit fly (Drosophila melanogaster), Chinese Hamster
Predicted to work with
Arabidopsis thaliana, Caenorhabditis elegans, Schizosaccharomyces pombe
Immunogen
Synthetic peptide derived from within residues 100 to the C-terminus of Human Histone H3.
(Peptide available as ab12149.)
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 7.4
Concentration
Concentration information loading...
Purity
Protein G purified
Primary antibody notes
This antibody is raised using the same immunogen as our popular anti-Histone H3 rabbit polyclonal - ab1791
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Unmodified
Applications
Show applications key
Our Abpromise guarantee covers the use of ab12079 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ChIP: Use 2-4 µg for 25 µg of chromatin. (Every new batch of this antibody is tested at Abcam in ChIP.)
IP: Use at an assay dependent dilution.
ChIP/Chip: Use at an assay dependent dilution.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Histone H3 peptide (ab12149).
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 histone antibody
- H3 histone family member E pseudogene antibody
- H3 histone family member E pseudogene antibody
see all
Database links
- Entrez Gene: 176359 Caenorhabditis elegans
- 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
- Entrez Gene: 15079 Mouse
- Entrez Gene: 15080 Mouse
see all
Anti-Histone H3 antibody - ChIP Grade images:
ChIP - anti-Histone H3 antibody - ChIP Grade (ab12079)
Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab12079 (blue), and 20 µl 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 - anti-Histone H3 antibody - ChIP Grade (ab12079)
Anti-Histone H3 antibody - ChIP Grade (ab12079) at 0.5 µg/ml + HeLa Cell Histone prep at 5 µg
Secondary
Rabbit polyclonal Secondary Antibody to Goat IgG - H&L (HRP) (ab6741) at 1/5000 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?)
Western blot - Histone H3 antibody (ab12079)
All lanes : Anti-Histone H3 antibody - ChIP Grade (ab12079) at 1 µg/ml
Lane 1 : Chicken S1:29 Cell lysate at 10 µg
Lane 2 : CHO Cell lysate at 10 µg
Lane 3 : Dog MDCK Cell lysate at 10 µg
Lane 4 : Mouse 3T3 Whole Cell Lysate
Lane 5 : Xenopus embryo Cell lysate at 10 µg
Secondary
Alexa Fluor 680 Rabbit anti-Goat IgG 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?)
References for Anti-Histone H3 antibody - ChIP Grade (ab12079)
This product has been referenced in:
- Wang Yet al. MicroRNA-138 modulates DNA damage response by repressing histone H2AX expression. Mol Cancer Res 9:1100-11 (2011). WB; Human.Read more (PubMed: 21693595) »
- Ricardi MMet al. Protocol: fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato. Plant Methods 6:11 (2010). ChIP; Plants.Read more (PubMed: 20380723) »
See all 3 publications for this product
Publishing research using ab12079? 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"