Anti-Histone H3 (di methyl K9)抗体[Y49] - ChIP Grade (ab32521)
- 数据表
- 具体的参考文献 (1)
- 实验方案
概述
- 产品名称Anti-Histone H3 (di methyl K9)抗体[Y49] - ChIP Grade
参阅全部 Histone H3 一抗 - 描述兔单克隆抗体[Y49] to Histone H3 (di methyl K9) - ChIP Grade
- 特异性The antibody only detects Histone H3 dimethylated on Lysine 9.
- 经测试应用适用于:
ICC/IF, WB, Flow Cyt, ChIPmore details
不适用于: IHC or IP - 种属反应性与反应: Mouse, Rat, Human
- 免疫原
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Histone H3 aa 1-100 (di methyl K9).
- 阳性对照
- WB: HeLa cell lysate. ICC/IF: HeLa cells. FC: HeLa cells.
- 常规说明
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
A trial size is available to purchase for this antibody.
性能
- 形式Liquid
- 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
- 存储溶液PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
- 纯度Tissue culture supernatant
- 克隆单克隆
- 克隆编号Y49
- 同种型IgG
- 研究领域
相关产品
- Compatible Secondaries
应用
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.
| 应用 | Ab评论 | 说明 |
|---|---|---|
| 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. |
靶标
- 功能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.
- 序列相似性Belongs to the histone H3 family.
- 发展阶段Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
- 翻译后修饰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. - 细胞定位Nucleus. Chromosome.
- Information by UniProt
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数据库链接
- Entrez Gene: 8290 Human
- 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 -
别名
- HIST1 cluster, H3J antibody
- Histone gene cluster 1, H3 histone family, member E antibody
- Histone gene cluster 1, H3G antibody
see all
Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade 图像
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![Western blot - Histone H3 (di methyl K9) antibody [Y49] (ab32521)](http://www.abcam.cn/ps/products/32/ab32521/Images/H3_WB.jpg)
Western blot - Histone H3 (di methyl K9) antibody [Y49] (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)](http://www.abcam.cn/ps/products/32/ab32521/Images/ab32521-265745-ab32521-Hela500.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)](http://www.abcam.cn/ps/products/32/ab32521/Images/ab32521-5-ab32521FC1.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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![ChIP - Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)](http://www.abcam.cn/ps/products/32/ab32521/Images/ab32521-254279-ChIPab325211.jpg)
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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![Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] (ab32521)](http://www.abcam.cn/ps/products/32/ab32521/Images/Histone-H3-Primary-antibodies-ab32521-3.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K9) antibody [Y49] (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.
Anti-Histone H3 (di methyl K9) antibody [Y49] - ChIP Grade (ab32521)参考文献
This product has been referenced in:
- Dalvoy Vasudevarao M et al. Inhibition of acetyltransferase alters different histone modifications: probed by small molecule inhibitor plumbagin. J Biochem 152:453-62 (2012). Read more (PubMed: 22923743) »
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"