Anti-Histone H3 (acetyl K9)抗体- ChIP Grade (ab10812)
- 数据表
- 具体的参考文献 (75)
- 实验方案
概述
- 产品名称Anti-Histone H3 (acetyl K9)抗体- ChIP Grade
参阅全部 Histone H3 一抗 - 描述兔多克隆抗体to Histone H3 (acetyl K9) - ChIP Grade
- 特异性Specific for acetyl K9, but cross-reacts slightly with K27.
- 经测试应用适用于: IHC-Fr, IHC-P, ChIP, IP, WB, CHIPseq, ICC/IF, Flow Cytmore details
- 种属反应性与反应: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish, Common marmoset, Rice
预测可用于: a wide range of other species
- 免疫原
Synthetic peptide corresponding to Human Histone H3 aa 1-100 (N terminal) (acetyl K9) conjugated to Keyhole Limpet Haemocyanin (KLH).
(Peptide available asab16635) - 阳性对照
- Calf Thymus Histone Preparation; Hela whole cell extract
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 or -80°C. Avoid freeze / thaw cycle.
- 存储溶液Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS. pH 7.4 -
Concentration information loading... - 纯度Immunogen affinity purified
- 克隆多克隆
- 同种型IgG
- 研究领域
相关产品
- ChIP Related Products
- Compatible Secondaries
- Corresponding Unmodified Peptide
- Immunizing Peptide (Blocking)
- Related Products
应用
Our Abpromise guarantee covers the use of ab10812 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 |
|---|---|---|
| IHC-Fr | 1/500. | |
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ChIP | Use 2-4 µg for 25 µg of chromatin. | |
| IP | Use at 20 µg/mg of lysate. | |
| WB | 1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). | |
| CHIPseq | Use at an assay dependent concentration. PubMed: 22196736 | |
| ICC/IF | Use a concentration of 1 µg/ml. | |
| Flow Cyt | Use at an assay dependent concentration. ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
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: 107131750 Cow
- Entrez Gene: 517139 Cow
- Entrez Gene: 616800 Cow
- Entrez Gene: 616819 Cow
- Entrez Gene: 788250 Cow
- Entrez Gene: 8290 Human
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
see all -
别名
- H3.3A antibody
- HIST1 cluster, H3E antibody
- H3 histone family, member A antibody
see all
Anti-Histone H3 (acetyl K9) antibody - ChIP Grade 图像
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Western blot - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812) at 1 µg/ml
Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (unmodified ) peptide (ab2903) at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (acetyl K9) peptide (ab16635) at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - acetyl K14 at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (acetyl K18) peptide (ab24003) at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (acetyl K27) peptide (ab24404) at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate withHuman Histone H3 (acetyl K23) peptide (ab48359) at 0.5 µg/ml
Lysates/proteins at 0.5 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Exposure time : 30 seconds -
ChIP - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)Chromatin was prepared from Hela 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 ab10812 (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). Primers and probes are located in the first kb of the transcribed region.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)IHC image of Histone H3 (acetyl K9) staining in human breast carcinoma FFPE section, performed on a BondTM system using the standard protocol F. 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 ab10812, 1µg/ml, for 8 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. -
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)ICC/IF image of ab10812 stained HeLa cells. The cells were 100% Methanol fixed (5 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab10812, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% Methanol fixed (5 min) Hek293, HepG2, and MCF-7 cells at 5µg/ml. -
Western blot - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)Image courtesy of an anonymous Abreview.All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812) at 1/1000 dilution
Lane 1 : Tissue lysate prepared from rat dorsal spinal cord at 5 µg
Lane 2 : Tissue lysate prepared from rat dorsal spinal cord at 5 µg
Lane 3 : Tissue lysate prepared from rat dorsal spinal cord at 8 µg
Lane 4 : Tissue lysate prepared from rat dorsal spinal cord at 8 µg
Lane 5 : Tissue lysate prepared from rat dorsal spinal cord at 8 µg withHuman Histone H3 (acetyl K9) peptide (ab16635) Each sample is from a different extract.
Secondary
HRP conjugated rabbit polyclonal
Developed using the ECL technique
Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)
Exposure time : 2 minutesImage courtesy of an anonymous Abreview.
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Immunohistochemistry (Frozen sections) - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)This image is courtesy of an Abreview submitted by Annabelle Kleinab10812 staining Histone H3 (acetyl K9) in Mouse thyroid tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Tween 20 and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/200 in 10% serum in PBS) for 1 hour at 24°C. An Alexa Fluor® 555-conjugated Goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)ab10812 staining histone H3 (acetyl K9) in A549 cells treated with scriptaid (ab120883), by ICC/IF. Increase in histone H3 (acetyl K9) expression correlates with increased concentration of scriptaid, as described in literature.
The cells were incubated at 37°C for 24 hour in media containing different concentrations of ab120883 (scriptaid) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab10812 (0.1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A anti-rabbit DyLight 488 secondary antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
Immunocytochemistry - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)This image is courtesy of Darin McDonald, Cross Cancer InstituteSK-N-SH cells fixed with 4% paraformaldehyde, permeabilized in 0.5% Triton X-100 and incubated for 1 hour with ab10812 (1/1000). ab10812 clearly stained the nucleus (red). The cells were counterstained with DAPI (blue). 100x magnification.
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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)This image was kindly supplied by Ms Vedrana Vicic by Abreviewab10812 staining Histone H3 (acetyl K9) in the root tips of Arabidopsis thaliana by Immunocytochemistry/ Immunofluorescence. Cells were fixed in 2% paraformaldehyde for 30 minutes at room temperature. Blocking and permeabilization was carried with 4% BSA solution containing 0,5% Triton X-100 in PBS at room temperature for 45 minutes. Slides were washed in PBS and incubated with primary antibody at a 1/200 dilution in 1% PBS for 1 hour at 37°C. Slides were washed in PBS and incubated with the secondary antibody ab6639 (Goat anti-rabbit Cy3 ® (H&L)) at a 1/500 dilution in 1% BSA in PBS for 1 hour at 37°C. Slides were counterstained with DAPI (2µg/mL) for 10 minutes at room temperature.Left image: DAPI stained interphase nucleus with prominent chromocentersMiddle image: Distribution of Histone H3 (acetyl K9) in the nucleus (arrows indicate absence of signal from chromocenters)Right image: Merged image
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Flow Cytometry - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)This image is courtesy of an Abreview submitted by Prof Albrecht Müllerab10812 staining mouse embryonic stem cells by flow cytometry (gated on all living cells). The cells were incuabted with the antibody at 0.25ug/1.5 x 105 cells in a permeabilization buffer. A PE conjugated goat anti-rabbit antibody was used as the secondary.
Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab10812)参考文献
This product has been referenced in:
- Ye Y et al. Chromatin remodeling during in vivo neural stem cells differentiating to neurons in early Drosophila embryos. Cell Death Differ 24:409-420 (2017). ChIP ; Drosophila melanogaster . Read more (PubMed: 27858939) »
- Yang G et al. The histone H3K9 methyltransferase SUV39H links SIRT1 repression to myocardial infarction. Nat Commun 8:14941 (2017). ChIP . Read more (PubMed: 28361889) »
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