![ChIP - Histone H3 (di methyl K4) antibody [Y47] (ab32356) image](#ab32356_1.jpg)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] (ab32356) image](#ab32356ICC.bmp)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356) image](#Histone-H3-Primary-antibodies-ab32356-3.jpg)
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Product name
Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade
See all Histone H3 products (41) ...
Description
Rabbit monoclonal [Y47] to Histone H3 (di methyl K4) - ChIP Grade
Specificity
This antibody only detects Histone H3 dimethylated on Lysine 4.
Tested applications
ChIP, CHIPseq, WB, IHC-P, ICC/IF, IPmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Predicted to work with
Sheep, Chicken, Cow, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Zebrafish, all Mammals, Rice
Immunogen
A synthetic (Dimethyl K) peptide corresponding to residues surrounding Lys4 of Histone H3.
Positive control
WB: HeLa cell lysate. IF: HeLa cells. IHC-P: Breast carcinoma.
General notes
Produced under U.S. Patent No. 5,675,063.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
Concentration
Concentration information loading...
Clonality
Monoclonal
Clone number
Y47
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 ab32356 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ChIP: Use at an assay dependent dilution. Use 8µl for 25µg of chromatin.
CHIPseq: Use at an assay dependent dilution. PubMed: 19581485
WB: 1/1000 - 1/10000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
IHC-P: 1/100 - 1/250.
ICC/IF: 1/250.
IP: Use at an assay dependent concentration. PubMed: 22086061
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 (di methyl K4) antibody [Y47] - ChIP Grade images:
ChIP - Histone H3 (di methyl K4) antibody [Y47] (ab32356)
![ChIP - Histone H3 (di methyl K4) antibody [Y47] (ab32356)](/ps/datasheet/Images/32/ab32356/ab32356_1.jpg)
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, 8µl of ab32356 (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.
Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] (ab32356)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] (ab32356)](/ps/datasheet/Images/32/ab32356/ab32356ICC.bmp)
Ab32356, at a 1/250 dilution, staining Histone H3 in HeLa cells by Immunofluorescence.
Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356)](/ps/datasheet/images/32/ab32356/Histone-H3-Primary-antibodies-ab32356-3.jpg)
ab32356 (1/200) staining Histone H3 (di methyl K4) in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus. For further experimental details please refer to Abreview.
Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356)
![Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356)](/ps/datasheet/images/32/ab32356/Histone-H3-Primary-antibodies-ab32356-4.jpg)
ICC/IF image of ab32356 stained HepG2 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 (ab32356 , 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) 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.
Western blot - Histone H3 (di methyl K4) antibody [Y47] (ab32356)
![Western blot - Histone H3 (di methyl K4) antibody [Y47] (ab32356)](/ps/datasheet/Images/32/ab32356/ab32356WB.bmp)
All lanes : Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356) at 1/20000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : Non methylated recombinant Histone H3
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Immunohistochemistry (Paraffin-embedded sections) - Histone H3 (di methyl K4) antibody [Y47] (ab32356)
![Immunohistochemistry (Paraffin-embedded sections) - Histone H3 (di methyl K4) antibody [Y47] (ab32356)](/ps/datasheet/Images/32/ab32356/ab32356IHCP.bmp)
Ab32356, at a 1/100 dilution, staining Histone H3 in paraffin embedded breast carcinoma tissue sections by Immunohistochemistry.
References for Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade (ab32356)
This product has been referenced in:
- Patel M et al. Tumor-specific retargeting of an oncogenic transcription factor chimera results in dysregulation of chromatin and transcription. Genome Res 22:259-70 (2012). IP; Human.Read more (PubMed: 22086061) »
- Tiwari VK et al. Target genes of Topoisomerase IIß regulate neuronal survival and are defined by their chromatin state. Proc Natl Acad Sci U S A 109:E934-43 (2012). ChIP.Read more (PubMed: 22474351) »
See all 11 publications for this product
Publishing research using ab32356? Please let us know so that we can cite the reference in this datasheet
![Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade for ChIP in Rice (32356)](/ps/Reviews/Images/ab9291_46076.jpg)
![Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade for Immunocytochemistry/ Immunofluorescence in Human (32356)](/ps/datasheet/images/32/ab32356/Histone-H3-Primary-antibodies-ab32356-2.jpg)
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"