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Product name
Anti-Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade
See all Histone H3 products (41) ...
Description
Rabbit polyclonal to Histone H3 (asymmetric di methyl R17) - ChIP Grade
Specificity
Peptide competition experiments confirmed that the antibody recognises specifically methylated R17 in H3 and not unmethylated H3 or methylated R3 in H4 (see figure 1). In whole cell extract the antibody recognises specifically only the methylated histone H3 protein band (see figure 2) Further, the antibody doesn't crossreact with the C-terminal methylation sites of CARM1 in histone H3. In IHC on paraffin-embedded sections of human tonsil, the antibody shows nuclear staining across most nuclei.
Tested applications
ChIP/Chip, IHC-P, IF, ICC/IF, IHC-P, ChIP, Dot Blot, IP, WBmore details
Cross reactivity
Reacts with
Chicken, Cow, Human, Fruit fly (Drosophila melanogaster), Toxoplasma gondii
Predicted to work with
all Mammals
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, di methylated at R17.
(Peptide available as ab169 35.)
Positive control
This antibody gave a positive signal in HeLa Histone Preparation Nuclear whole cell lysate.
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: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Primary antibody notes
The nuclear hormone receptor co-activator CARM1 has the potential to methylate histone H3 at arginine residues in vitro. The methyltransferase activity of CARM1 is necessary for its co-activator functions in transient transfection assays. However, the role of this methyltransferase in vivo is unclear, given that methylation of arginines is not easily detectable on purified histones. This antibody recognizes methylated arginine 17 (R17) of histone H3, the major site of methylation by CARM1. Bauer et al (2001) have shown by using this antibody that methylated R17 exists in vivo. Chromatin immunoprecipitation analysis shows that R17 methylation on histone H3 is dramatically upregulated when the estrogen receptor-regulated pS2 gene is stimulated by estradiol and TPA. Coincident with the appearance of methylated R17, the CARM1 methyltransferase is found associated with the histones on the pS2 gene. Together these results demonstrate that the CARM1 methyltransferase is recruited to an active promoter and that CARM1-mediated methylation of histone H3 at R17 takes place in vivo during this active state.
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
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- Histone H3 (di methyl R17) antibody - ChIP Grade (ab8284)
(enlarge)
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Immunocytochemistry/ Immunofluorescence - Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284)
(enlarge)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284)
(enlarge)
Applications
Show applications key
Our Abpromise guarantee covers the use of ab8284 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ChIP/Chip: Use at an assay dependent dilution.
IHC-P: Use at an assay dependent dilution.
IF
IF: 1/1000.IF: 1/1000.
ICC/IF: Use at an assay dependent concentration.
IHC-P: Use at an assay dependent concentration.
ChIP: Use at an assay dependent dilution.
Dot: Use at an assay dependent dilution.
IP: Use at an assay dependent dilution.
WB: 1/1000 - 1/2000.
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 accession
P68431
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 (asymmetric di methyl R17) antibody - ChIP Grade images:
- Histone H3 (di methyl R17) antibody - ChIP Grade (ab8284)
Total U2OS cell extract was western blotted using the anti-Me-R17H3 antibody. The asterisk indicates methylated histone H3. The left panel shows presence of core histones (indicated on the left) by Coomassie Blue staining. Molecular weights are indicated on the right.
Taken from Bauer et al, (2001).
Immunocytochemistry/ Immunofluorescence - Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284)
ab8284 was used in immunohistochemistry with paraffin embedded sections of human tonsil, using DAB as a chromogen (brown). Counterstaining of nuclei was performed with haemotoxylin (blue).
Staining is seen confined to the nucleus.
Dot Blot - Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284)
A dot blot was performed using unmodified peptide (lane 1), Histone H3 mono methyl R17 peptide (lane 2), Histone H3 asymmetric di methyl R17 peptide (lane 3) and Histone H3 symmetric di methyl R17 peptide (lane 4). The dot blot indicates that ab8284 is specific to Histone H3 asymmetric di methyl R17.
Western blot - Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284)
All lanes : Anti-Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284) at 1 µg/ml
Lane 1 : HeLa Histone Preparation Nuclear Lysate
Lane 2 : HeLa Histone Preparation Nuclear Lysate with
Lane 3 : HeLa Histone Preparation Nuclear Lysate with
Lane 4 : HeLa Histone Preparation Nuclear Lysate with
Lysates/proteins at 2.5 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 15.4 kDa
Observed band size : 16 kDa (why is the actual band size different from the predicted?)
Additional bands at : 60 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 2 minutes
References for Anti-Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284)
This product has been referenced in:
- Bauer UM et al. Methylation at arginine 17 of histone H3 is linked to gene activation. EMBO Rep 3:39-44 (2002).Read more (PubMed: 11751582) »
- Wu J & Xu W Histone H3R17me2a mark recruits human RNA Polymerase-Associated Factor 1 Complex to activate transcription. Proc Natl Acad Sci U S A 109:5675-80 (2012).Read more (PubMed: 22451921) »
See all 18 publications for this product
Publishing research using ab8284? 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"