製品名Anti-Histone H3 (acetyl K18) antibody - ChIP Grade
Histone H3 一次抗体 製品一覧
製品の詳細Rabbit polyclonal to Histone H3 (acetyl K18) - ChIP Grade
特異性Specificity for acetyl K18 has been validated by blocking peptide/antibody incubation followed by immunostaining western blots.
アプリケーション適用あり: WB, PepArr, CHIPseq, IHC-P, ICC/IF, ChIP/Chip, ChIPmore details
種交差性交差種: Mouse, Rat, Cow, Human, Saccharomyces cerevisiae, Arabidopsis thaliana, Drosophila melanogaster, Fungi
Synthetic peptide corresponding to Histone H3 aa 1-100 (acetyl K18) conjugated to Keyhole Limpet Haemocyanin (KLH).
(Peptide available as
- This antibody gave a positive signal in HeLa Histone Preparation Nuclear Lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal skin.
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.
Concentration information loading...
精製度Immunogen affinity purified
ChIP Related Products
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab1191 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 17 kDa.|
|PepArr||Use a concentration of 0.2 - 0.02 µg/ml.|
|CHIPseq||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|ChIP/Chip||Use at an assay dependent concentration.|
|ChIP||Use 2µg for 106 cells.|
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 U2OS 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 ab1191 (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 (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
All batches of ab1191 are tested in Peptide Array against peptides to different Histone H3 and H4 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 - acetyl K18 peptide (ab24003), indicating that this antibody specifically recognises the Histone H3 - acetyl K18 modification.
ab17163 - Histone H3 - unmodified
ab16635 - Histone H3 - acetyl K9
ab15591 - Histone H3 - acetyl K14
ab24003 - Histone H3 - acetyl K18
ab48359 - Histone H3 - acetyl K23
ab24404 - Histone H3 - acetyl K27
ab41409 - Histone H3 - acetyl K36
ab15662 - Histone H4 - acetyl K12
All lanes : Anti-Histone H3 (acetyl K18) antibody - ChIP Grade (ab1191) at 1 µg/ml
Lane 1 : HeLa Histone Preparation Nuclear Lysate
Lane 2 : HeLa Histone Preparation Nuclear Lysate with Histone H3 peptide - unmodified R17 at 0.5 µg/ml
Lane 3 : HeLa Histone Preparation Nuclear Lysate with
Human Histone H3 (acetyl K9) peptide (ab16635) at 0.5 µg/ml
Lane 4 : HeLa Histone Preparation Nuclear Lysate with Histone H3 peptide - acetyl K14 at 0.5 µg/ml
Lane 5 : HeLa Histone Preparation Nuclear Lysate with
Human Histone H3 (acetyl K18) peptide (ab24003) at 0.5 µg/ml
Lane 6 : HeLa Histone Preparation Nuclear Lysate with
Human Histone H3 (acetyl K23) peptide (ab48359) at 0.5 µg/ml
Lane 7 : HeLa Histone Preparation Nuclear Lysate with
Human Histone H3 (acetyl K27) peptide (ab24404) at 0.5 µg/ml
Lane 8 : HeLa Histone Preparation Nuclear Lysate with Histone H3 peptide - acetyl K36 at 0.5 µg/ml
Lane 9 : HeLa Histone Preparation Nuclear Lysate with
Human Histone H4 (acetyl K12) peptide (ab15662) at 0.5 µg/ml
Lysates/proteins at 2.5 µg per lane.
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 15.4 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 1 minute
L929, 3T3 and HeLa cells fixed with 2% paraformaldehyde in PBS were incubated for 5 min in 0.2% Triton X-100 in PBS before a blocking step in 5% BSA, 0.1% Tween 20 in PBS (blocking buffer). Ac K18-H3 (ab1191) antibody was added diluted at 1/500 in blocking buffer for a 45 min incubation at room temperature. For visualization, fluorescein isothiocyanate (FITC) anti rabbit secondary antibody was used diluted at 1/800. DAPI (Sigma) in 0.1% Tween 20 in PBS was used in the final washing steps to stain DNA. Vectashield (Vector Laboratories Inc.) was used as mounting medium.
The ac K18-H3 antibody revealed a granular staining throughout the nucleoplasm, excluding regions that show an intense DAPI staining (mostly pericentric heterochromatin in mice cells).
IHC image of Histone H3 (acetyl K18) staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab1191, 1µg/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.
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.
ICC/IF image of ab1191 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 (ab1191, 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 MCF7 cells at 1µg/ml, and in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 1µg/ml.
This product has been referenced in:
- Fellows R et al. Microbiota derived short chain fatty acids promote histone crotonylation in the colon through histone deacetylases. Nat Commun 9:105 (2018). Read more (PubMed: 29317660) »
- Hsu KW et al. The Application of Non-Invasive Apoptosis Detection Sensor (NIADS) on Histone Deacetylation Inhibitor (HDACi)-Induced Breast Cancer Cell Death. Int J Mol Sci 19:N/A (2018). Read more (PubMed: 29393914) »