Anti-Histone H3 (di methyl K36) 抗体 [EPR16994(2)] - ChIP Grade (ab176921)
![ChIP - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-9-AntiHistone-H3-di-methyl-K36-antibody-EPR169942--ChIP-GradeChIP.jpg "ChIP - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16994(2)] to Histone H3 (di methyl K36) - ChIP Grade
- Suitable for: PepArr, ICC/IF, ChIP, IHC-P, WB
- Reacts with: Mouse, Rat, Human
製品の概要
-
製品名
Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade
Histone H3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR16994(2)] to Histone H3 (di methyl K36) - ChIP Grade -
由来種
Rabbit -
アプリケーション
適用あり: PepArr, ICC/IF, ChIP, IHC-P, WBmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Histone H3 aa 1-100 (di methyl K36). The exact sequence is proprietary.
Database link: P68431 -
ポジティブ･コントロール
- WB: HeLa and NIH/3T3 whole cell lysates. IHC-P: Human stomach, mouse testis and rat spleen tissues. ICC/IF: HeLa and NIH/3T3 cells. ChIP: Chromatin from HeLa cells.
-
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
-
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 long term. Avoid freeze / thaw cycle. -
バッファー
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
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
EPR16994(2) -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
アプリケーション
Our Abpromise guarantee covers the use of ab176921 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 |
|---|---|---|
| PepArr | Use at an assay dependent concentration. | |
| ICC/IF | 1/500. | |
| ChIP | Use 2 µg for 25 µg of chromatin. | |
| IHC-P | 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| WB | 1/5000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa). |
ターゲット情報
-
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
-
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
-
ChIP - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab176921 (red), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
-
![Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-255669-anti-histone-h3-di-methyl-k36-antibody-epr169942-chip-grade-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Histone H3 (di methyl K36) with ab176921 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab176921 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution. -
![Peptide Array - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-261515-anti-histone-h3-di-methyl-k36-antibody-epr169942-chip-grade-peptide-array.jpg)
Peptide Array - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)ab176921 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here. -
![Western blot - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-255677-anti-histone-h3-di-methyl-k36-antibody-epr169942-chip-grade-western-blot.jpg)
Western blot - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)All lanes : Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921) at 1/5000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Predicted band size: 15 kDa
Observed band size: 15 kDa
Exposure time: 4 secondsBlocking/Dilution buffer: 5% BSA/TBST.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-255675-anti-histone-h3-di-methyl-k36-antibody-epr169942-chip-grade-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Histone H3 (di methyl K36) with ab176921 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human stomach is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-255672-anti-histone-h3-di-methyl-k36-antibody-epr169942-chip-grade-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Histone H3 (di methyl K36) with ab176921 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse testis is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-255671-anti-histone-h3-di-methyl-k36-antibody-epr169942-chip-grade-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Histone H3 (di methyl K36) with ab176921 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat spleen is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-255670-anti-histone-h3-di-methyl-k36-antibody-epr169942-chip-grade-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Histone H3 (di methyl K36) with ab176921 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: ab176921 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution. -
![Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)](https://www.abcam.co.jp/ps/products/176/ab176921/Images/ab176921-10-benefits-of-recombinant-antibodies.png)
Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
プロトコール
データシートおよび資料
Certificate of Compliance
参考文献 (2)
ab176921 は 2 報の論文で使用されています。
- Zhou R et al. Characterization of H3 methylation in regulating oocyte development in cyprinid fish. Sci China Life Sci N/A:N/A (2018). PubMed: 30443860
- Streubel G et al. The H3K36me2 Methyltransferase Nsd1 Demarcates PRC2-Mediated H3K27me2 and H3K27me3 Domains in Embryonic Stem Cells. Mol Cell 70:371-379.e5 (2018). PubMed: 29606589
