Anti-Histone H3 (di methyl K9) 抗体 [Y49] - ChIP Grade (ab32521)
![ChIP - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)](https://www.abcam.co.jp/ps/products/32/ab32521/Images/ab32521-254279-anti-histone-h3-di-methyl-k9-antibody-y49-chip-grade-chip.jpg "ChIP - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y49] to Histone H3 (di methyl K9) - ChIP Grade
- Suitable for: ICC/IF, WB, Flow Cyt, ChIP
- Reacts with: Mouse, Rat, Human
製品の概要
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製品名
Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade
Histone H3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [Y49] to Histone H3 (di methyl K9) - ChIP Grade -
由来種
Rabbit -
特異性
The antibody only detects Histone H3 dimethylated on Lysine 9.
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アプリケーション
適用あり: ICC/IF, WB, Flow Cyt, ChIPmore details
適用なし: IHC or IP -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Histone H3 aa 1-100 (di methyl K9). The exact sequence is proprietary.
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ポジティブ･コントロール
- WB: HeLa cell lysate. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. ChIP: Chromatin prepared from HeLa cells.
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
Y49 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
アプリケーション
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.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| ICC/IF | 1/500. | |
| WB | 1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). | |
| Flow Cyt | 1/220. | |
| ChIP | Use 5 µg for 25 µg of chromatin. |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the histone H3 family. -
発生段階
Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. -
翻訳後修飾
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. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
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参照データベース
- 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
- Entrez Gene: 8357 Human
see all -
別名
- H3 histone family member E pseudogene antibody
- H3 histone family, member A antibody
- H3/A antibody
see all
画像
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ChIP - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab32521 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
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![Western blot - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)](https://www.abcam.co.jp/ps/products/32/ab32521/Images/ab32521-45100-anti-histone-h3-di-methyl-k9-antibody-y49-chip-grade-western-blot.jpg)
Western blot - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)All lanes : Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (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] - ChIP Grade (ab32521)](https://www.abcam.co.jp/ps/products/32/ab32521/Images/ab32521-265745-anti-histone-h3-di-methyl-k9-antibody-y49-chip-grade-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Histone H3 (di methyl K9) with purified ab32521 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
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![Flow Cytometry - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)](https://www.abcam.co.jp/ps/products/32/ab32521/Images/ab32521-270691-anti-histone-h3-di-methyl-k9-antibody-y49-chip-grade-flow-cytometry.jpg)
Flow Cytometry - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Histone H3 (di methyl K9) with purified ab32521 at 1/220 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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![Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)](https://www.abcam.co.jp/ps/products/32/ab32521/Images/ab32521-45099-anti-histone-h3-di-methyl-k9-antibody-y49-chip-grade-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canadaab32521 (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.
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![Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)](https://www.abcam.co.jp/ps/products/32/ab32521/Images/ab32521-6-benefits-of-recombinant-antibodies.png)
Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
参考文献 (12)
ab32521 は 12 報の論文で使用されています。
- Casciello F et al. G9a-mediated repression of CDH10 in hypoxia enhances breast tumour cell motility and associates with poor survival outcome. Theranostics 10:4515-4529 (2020). PubMed: 32292512
- Liu N et al. MicroRNA-181 regulates the development of Ossification of Posterior longitudinal ligament via Epigenetic Modulation by targeting PBX1. Theranostics 10:7492-7509 (2020). PubMed: 32685001
- Kim GH et al. IDH1R132H Causes Resistance to HDAC Inhibitors by Increasing NANOG in Glioblastoma Cells. Int J Mol Sci 20:N/A (2019). PubMed: 31151327
- Lu KL et al. Transgenerational dynamics of rDNA copy number inDrosophilamale germline stem cells. Elife 7:N/A (2018). PubMed: 29436367
- Wang C et al. MeCP2-mediated epigenetic regulation in senescent endothelial progenitor cells. Stem Cell Res Ther 9:87 (2018). PubMed: 29615114
