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Product name
Anti-Histone H3 (mono methyl R2) antibody - ChIP Grade
See all Histone H3 products (41) ...
Description
Rabbit polyclonal to Histone H3 (mono methyl R2) - ChIP Grade
Tested applications
ChIP/Chip, IHC-P, PepArr, WB, ChIPmore details
Cross reactivity
Reacts with
Mouse, Cow, Human, Saccharomyces cerevisiae
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 to the C-terminus Histone H3, mono methylated at R2.
Positive control
Calf thymus histone, C3H10T1/2 and 293 lysates. In ICC/IF this antibody gave a positive result in HeLa cell line.
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
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> ChIP'ing antibodies >> ChIP'ing antibodies
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Applications
Show applications key
Our Abpromise guarantee covers the use of ab15584 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: 1/80. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
PepArr: Use a concentration of 0.2 - 2 µg/ml.
WB: Use a concentration of 0.5 - 2 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
ChIP: Use 2-4 µg for 25 µg of chromatin.
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: 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
- Entrez Gene: 8356 Human
see all
Anti-Histone H3 (mono methyl R2) antibody - ChIP Grade images:
ChIP - Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584)
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 ab15584 (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). Primers and probes are located in the first kb of the transcribed region.
Peptide Array - Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584)
All batches of ab15584 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - mono methyl R2 peptide (ab1775), indicating that this antibody specifically recognises the Histone H3 - mono methyl R2 modification.
1. ab1775 – Histone H3 - mono methyl R2 peptide
2. ab38488 – Histone H3 - symmetric di methyl R2
3. ab2853 – Histone H3 - asymmetric di methyl R2
4. ab17566 – Histone H3 - non-modified
5. ab33971 – Histone H3 - mono methyl R8
6. ab13845 – Histone H3 - mono methyl R17
7. ab16935 – Histone H3 - asymmetric di methyl R17
8. ab32948 – Histone H3 - symmetric di methyl R17
9. ab2854 – Histone H3 - asymmetric di methyl R26
10. ab38487 – Histone H3 - symmetric di methyl R26
Western blot - Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584)
Anti-Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584) at 1 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 17 kDa
Exposure time : 3 minutes
Western blot - Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584)
All lanes : Anti-Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584) at 0.5 µg/ml
Lane 1 : Calf thymus histone at 0.4 µg
Lane 2 : C3H10T1/2 histones at 0.4 µg
Lane 3 : 293 histones at 0.4 µg
Lane 4 : Yeast extracts from 1 x 106 cells
Predicted band size : 17 kDa
ab15584 specifically recognises mono-methylated Histone H3 at R2 in calf thymus (lane1), mouse (lane2) and human (lane3) histone preparations as well as in yeast whole cell extract (lane4).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584)
Image courtesy of Human Protein Atlas
ab15584 staining histone H3 mono methyl R2 in human testis, showing a distinct and strong nuclear staining pattern in ductus seminiferus and leydig cells. Paraffin embedded human skin tissue was incubated with ab15584 (1/80 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.
ab15584 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584)
ICC/IF image of ab15584 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab15584, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-Histone H3 (mono methyl R2) antibody - ChIP Grade (ab15584)
This product has been referenced in:
- Chignola F et al. The solution structure of the first PHD finger of autoimmune regulator in complex with non-modified histone H3 tail reveals the antagonistic role of H3R2 methylation. Nucleic Acids Res 37:2951-61 (2009). WB; Human.Read more (PubMed: 19293276) »
- Iberg AN et al. Arginine methylation of the histone h3 tail impedes effector binding. J Biol Chem 283:3006-10 (2008). IHC-P; Mouse.Read more (PubMed: 18077460) »
See all 4 publications for this product
Publishing research using ab15584? 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"