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Products:Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
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Read our guarantee »Anti-Histone H3 (di methyl K9) antibody [Y49]
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Rabbit monoclonal [Y49] to Histone H3 (di methyl K9)
The antibody only detects Histone H3 dimethylated on Lysine 9.
ICC/IF, ICC, WBmore details
Reacts with
Mouse, Rat, Human
Synthetic (Dimethyl-K)-peptide corresponding to residues surrounding Lys9 of Histone H3
HeLa cell lysate
Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
Tissue culture supernatant
Monoclonal
Y49
IgG
Epigenetics and Nuclear Signaling >> Histones >> H3 >> Methylated
Our Abpromise guarantee covers the use of ab32521 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC/IF: 1/500.
ICC: 1/100.
WB: 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
Is unsuitable for ,Flow Cyt,IHC or IP.
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.
Belongs to the histone H3 family.
Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
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.
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).
Western blot - Histone H3 (di methyl K9) antibody [Y49] (ab32521)
![Western blot - Histone H3 (di methyl K9) antibody [Y49] (ab32521)](/ps/datasheet/Images/32/ab32521/H3_WB.jpg)
All lanes : Anti-Histone H3 (di methyl K9) antibody [Y49] (ab32521) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : recombinant Histone H3
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] (ab32521)
![Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] (ab32521)](/ps/datasheet/images/32/ab32521/Histone-H3-Primary-antibodies-ab32521-3.jpg)
ab32521 (1/500) staining Histone H3 di-methyl K9 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). 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
ab32521 has not yet been referenced specifically in any publications.
Publishing research using ab32521? Please let us know so that we can cite the reference in this datasheet
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![Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] (ab32521)](/ps/datasheet/images/32/ab32521/Histone-H3-Primary-antibodies-ab32521-3.jpg)
ab32521 (1/500) staining Histone H3 di-methyl K9 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus (red). 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
![Anti-Histone H3 (di methyl K9) antibody [Y49] for Immunocytochemistry/ Immunofluorescence in Human (32521)](/ps/datasheet/images/32/ab32521/Histone-H3-Primary-antibodies-ab32521-2.jpg)
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