![Western blot - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147) image](#ab6147_1.jpg)
![ELISA - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147) image](#ab6147_2.gif)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147) image](#ab6147-IF-Im1.jpg)
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Product name
Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6147]
See all Histone H3 products (41) ...
Description
Mouse monoclonal [mAbcam 6147] to Histone H3 (tri methyl K27)
Specificity
By Western blot, this antibody is blocked strongly by tri methyl K27 peptide and weakly by di methyl K27 peptide. By ELISA, it is specific for tri methyl K27 peptide as shown (although it also binds very weakly with di methyl K27 peptide). The sequence which it reacts is found in all Mammals and a wide range of other species, including D. melanogaster, Arabidopsis, Chicken and Xenopus. The antibody will react with any of the above species where the modification is present. Reactivity is not certain in S. pombe and S. cerevisiae as the equivalent protein sequence differs slightly from species listed above.
Tested applications
ICC/IF, IHC-P, ELISA, WBmore details
Cross reactivity
Reacts with
Mouse, Cow, Human
Predicted to work with
Rat, Chicken, Xenopus laevis, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster)
Does not react with
Carp
Immunogen
Synthetic peptide derived from residues 1 - 100 of Human Histone H3, tri methylated at K27.
. Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising peptide. Clones were negatively screened against both the corresponding unmodified peptide and also against a peptide corresponding to tri methylated K9 of Histone H3.
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
IgG fraction
Clonality
Monoclonal
Clone number
mAbcam 6147
Isotype
IgG1
Light chain type
kappa
Research areas
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Applications
Show applications key
Our Abpromise guarantee covers the use of ab6147 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 a concentration of 1 µg/ml.
IHC-P: Use at an assay dependent concentration.
ELISA: Use a concentration of 1 µg/ml. PubMed: 17641388
WB: Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Histone H3 peptide - tri methyl K27 (ab1782).
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 accessionP68431
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Histone H3.1 antibody
- FLJ92264 antibody
- H3 histone antibody
see all
Database links
- Entrez Gene: 427887 Chicken
- Entrez Gene: 326601 Cow
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
- Entrez Gene: 8354 Human
- Entrez Gene: 8355 Human
see all
Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6147] images:
Western blot - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147)
![Western blot - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147)](/ps/datasheet/Images/6/ab6147/ab6147_1.jpg)
All lanes : Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147) at 5 µg/ml
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with
Lane 3 : Calf thymus histone lysate with
Lane 4 : Calf thymus histone lysate with
Lane 5 : Calf thymus histone lysate with
Lane 6 : Calf thymus histone lysate with
Lane 7 : Calf thymus histone lysate with
Lane 8 : Calf thymus histone lysate with
Lane 9 : Calf thymus histone lysate with
Lane 10 : Calf thymus histone lysate with
Lane 11 : Calf thymus histone lysate with
Secondary
Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6789) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 15 kDa
Exposure time : 2 minutes
ELISA - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147)
![ELISA - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147)](/ps/datasheet/Images/6/ab6147/ab6147_2.gif)
ELISA using ab6147.
The red line indicates binding to the tri methyl K27 peptide (ab1782). Weak binding (brown line) was seen to the di methyl K27 peptide (ab1781). Binding to the following peptides was not seen:
Unmodified K27 (ab2623),
mono methyl K27 (ab1780),
mono methyl K9 (ab1771),
di methyl K9 (ab1772),
tri methyl K9 (ab1773),
mono methyl K4 (ab1340),
di methyl K4 (ab7768),
tri methyl K4 (ab1342).
This indicates the specificity of ab6147 for tri methyl K27 of Histone H3.
Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147)](/ps/datasheet/Images/6/ab6147/ab6147-IF-Im1.jpg)
ICC/IF image of ab6147 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab6147, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
References for Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6147] (ab6147)
This product has been referenced in:
- Peach SE et al. Quantitative assessment of ChIP-grade antibodies directed against histone modifications reveals patterns of co-occurring marks on histone protein molecules. Mol Cell Proteomics : (2012).Read more (PubMed: 22442256) »
- Chaumeil J et al. Evolution from XIST-independent to XIST-controlled X-chromosome inactivation: epigenetic modifications in distantly related mammals. PLoS One 6:e19040 (2011). ICC/IF.Read more (PubMed: 21541345) »
See all 3 publications for this product
Publishing research using ab6147? Please let us know so that we can cite the reference in this datasheet
![Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6147] for Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) in Mouse (6147)](/ps/datasheet/images/6/ab6147/Histone-H3-Primary-antibodies-ab6147-4.jpg)
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