Key features and details
- Rabbit polyclonal to Histone H3 (mono methyl K79) - ChIP Grade
- Suitable for: ChIP, IP, WB, PepArr
- Reacts with: Mouse, Rat, Cow, Human
- Isotype: IgG
产品名称Anti-Histone H3 (mono methyl K79)抗体- ChIP Grade
参阅全部 Histone H3 一抗
描述兔多克隆抗体to Histone H3 (mono methyl K79) - ChIP Grade
特异性Reacts with Mono-methyl K79 of histone H3. Slight cross-reactivity to di-methyl K79.
经测试应用适用于: ChIP, IP, WB, PepArrmore details
种属反应性与反应: Mouse, Rat, Cow, Human
Synthetic peptide within Human Histone H3 aa 50 to the C-terminus (mono methyl K79) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as
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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.
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存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Preservative: 0.02% Sodium azide
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
Concentration information loading...
纯度Immunogen affinity purified
ChIP Related Products
Our Abpromise guarantee covers the use of ab2886 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use 2µg for 106 cells.|
|IP||Use at an assay dependent concentration.|
|WB||1/1 - 1/500. Detects a band of approximately 17 kDa.|
|PepArr||Use a concentration of 0.2 - 0.02 µg/ml.|
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.
- Information by UniProt
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Chromatin was prepared from U-2 OS (Human bone osteosarcoma epithelial cell line) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab2886 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR. Primers and probes are located in the first kb of the transcribed region.
Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886) at 1 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg
Human NFIB / NF1B2 peptide (ab95051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15.4 kDa
Additional bands at: 17 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 3 minutes
All batches of ab2886 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - mono methyl K79 peptide (ab4555), indicating that this antibody specifically recognises the Histone H3 - mono methyl K79 modification.
ab4558 - Histone H3 - unmodified
ab1771 - Histone H3 - di methyl K9
ab4555 - Histone H3 - mono methyl K79
ab4556 - Histone H3 - di methyl K79
ab4557 - Histone H3 - tri methyl K79
ab4560 - Histone H4 - di methyl K79
All lanes : Anti-Histone H3 (mono methyl K79) antibody - ChIP Grade (ab2886) at 1/300 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Testis (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15.4 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutes
Chromatin was prepared from whole cell lysate of normal rat liver and liver cancer cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. 5 µg of the primary antibody was used in 1/100 dilution and it was incubated with the sample for 16 hours at 4°C in a commercially available ChIP dilution buffer. The immunoprecipitated DNA was quantified by real time PCR. ChIP results show that the Histone H3 (mono methyl K79) and GAPDH genes are expressed in higher levels in liver cancer cells than in normal liver cells.
Histone H3 (mono methyl K79) was immunoprecipitated using 0.5mg HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 5µg of Rabbit polyclonal to Histone H3 (mono methyl K79) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2886.
Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
Band: 18kDa: Histone H3 (mono methyl K79).
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.
ab2886 被引用在 47 文献中.
- Liu C et al. CBP mediated DOT1L acetylation confers DOT1L stability and promotes cancer metastasis. Theranostics 10:1758-1776 (2020). PubMed: 32042335
- Nassa G et al. Inhibition of histone methyltransferase DOT1L silences ERa gene and blocks proliferation of antiestrogen-resistant breast cancer cells. Sci Adv 5:eaav5590 (2019). PubMed: 30775443
- Chory EJ et al. Nucleosome Turnover Regulates Histone Methylation Patterns over the Genome. Mol Cell 73:61-72.e3 (2019). PubMed: 30472189
- Lam KG et al. Cell-type-specific genomics reveals histone modification dynamics in mammalian meiosis. Nat Commun 10:3821 (2019). PubMed: 31444359
- Lu X et al. GLP-catalyzed H4K16me1 promotes 53BP1 recruitment to permit DNA damage repair and cell survival. Nucleic Acids Res 47:10977-10993 (2019). PubMed: 31612207