Key features and details
- Rabbit polyclonal to Histone H3 (mono methyl K4) - ChIP Grade
- Suitable for: ICC/IF, ChIP, WB, IHC-P
- Reacts with: Cow, Human
- Isotype: IgG
製品名Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade
Histone H3 一次抗体 製品一覧
製品の詳細Rabbit polyclonal to Histone H3 (mono methyl K4) - ChIP Grade
特異性Specific for mono-methylated Lysine 4 of histone H3. Does not recognise di- or tri-methyl Lysine 4 nor methylation at Lysine 9.
アプリケーション適用あり: ICC/IF, ChIP, WB, IHC-Pmore details
種交差性交差種: Cow, Human
交差が予測される動物種: Mouse, Pig, Saccharomyces cerevisiae, Tetrahymena, Xenopus laevis, Drosophila melanogaster, Plants, Mammals, Plasmodium falciparum, Xenopus tropicalis, Candida albicans
Synthetic peptide within Human Histone H3 aa 1-100 (mono methyl K4) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
(Peptide available as
- ChIP: Chromatin prepared from U-2 OS cells; Metastatic clear cell renal carcinoma cells (M1A). WB: Calf thymus histone lysate. ICC/IF: HeLa cells. IHC-P: Human colon tissue.
Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.
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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.
保存方法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.
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精製度Immunogen affinity purified
ChIP Related Products
- Human Histone H3 (tri methyl K4) peptide (ab1342)
- Human Histone H3 (mono methyl K9) peptide (ab1771)
- Human Histone H3 (di methyl K9) peptide (ab1772)
- Human Histone H3 (tri methyl K9) peptide (ab1773)
- Human Histone H3 (mono methyl K27) peptide (ab1780)
- Human Histone H3 (di methyl K27) peptide (ab1781)
- Human Histone H3 (tri methyl K27) peptide (ab1782)
- Human Histone H3 (unmodified) peptide (ab7228)
- Human Histone H3 (di methyl K4) peptide (ab7768)
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab8895 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.
Works better if cells are fixed with methanol.
|ChIP||Use 2 µg for 25 µg of chromatin.
We recommend Myo-D ChIP primer pair ab269261 as positive control.
|WB||1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).|
|IHC-P||Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
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.
- Information by UniProt
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Chromatin was prepared from U-2 OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab8895 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH and ALDOA (active) and MYO-D (inactive) promoters and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.
All lanes : Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895) at 1/500 dilution
Lane 1 : Calf thymus histone lysate
Lane 2 : Calf thymus histone lysate with
Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/ml
Lane 3 : Calf thymus histone lysate with
Human Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/ml
Lane 4 : Calf thymus histone lysate with
Human Histone H3 (tri methyl K4) peptide (ab1342) at 1 µg/ml
Lane 5 : Calf thymus histone lysate with
Human Histone H3 (mono methyl K9) peptide (ab1771) at 1 µg/ml
Lane 6 : Calf thymus histone lysate with
Human Histone H3 (mono methyl K27) peptide (ab1780) at 1 µg/ml
Lane 7 : Calf thymus histone lysate with
Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/ml
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 15 kDa
Exposure time: 2 minutes
ab8895 is specific for mono-methylated Lysine 4 of histone H3 and does not recognize di- or tri-methyl Lysine 4 nor methylation at Lysine 9. This is shown in lane 2 where the activity of the antibody is specifically blocked by the addition of the immunizing peptide (ab1340).
ab8895 staining Histone H3 (mono methyl K4) in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab8895 at 1µg/ml and ab7291 (anti beta Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat anti-rabbit AlexaFluor®488 secondary (ab150081) at 2 μg/ml (shown in green) and a goat anti-mouse AlexaFluor®594 secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
IHC image of ab8895 staining Histone H3 (mono methyl K4) in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8895, 0.5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ChIP analysis using ab8895 binding Histone H3 (mono methyl K4) in Metastatic clear cell renal carcinoma cells (M1A). Cells were cross-linked for 10 minutes with 1% formaldehyde. Samples were incubated at 1/100 dilution with primary antibody for 12 hours at 4°C.
Positive control:Metastatic clear cell renal carcinoma cells (M1A).
Negative Control:Parental clear cell renal carcinoma cells (786-O).
ab8895 は 794 報の論文で使用されています。
- Domingues AF et al. Loss of Kat2a enhances transcriptional noise and depletes acute myeloid leukemia stem-like cells. Elife 9:N/A (2020). PubMed: 31985402
- Kupai A et al. A Degenerate Peptide Library Approach to Reveal Sequence Determinants of Methyllysine-Driven Protein Interactions. Front Cell Dev Biol 8:241 (2020). PubMed: 32328492
- Glanzner WG et al. Histone Lysine Demethylases KDM5B and KDM5C Modulate Genome Activation and Stability in Porcine Embryos. Front Cell Dev Biol 8:151 (2020). PubMed: 32211412
- Yu YC et al. Transient DNMT3L Expression Reinforces Chromatin Surveillance to Halt Senescence Progression in Mouse Embryonic Fibroblast. Front Cell Dev Biol 8:103 (2020). PubMed: 32195249
- Lai Y et al. Coordinated regulation of infection-related morphogenesis by the KMT2-Cre1-Hyd4 regulatory pathway to facilitate fungal infection. Sci Adv 6:eaaz1659 (2020). PubMed: 32232158