Product nameAnti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade
See all Histone H3 primary antibodies
DescriptionRabbit polyclonal to Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) - ChIP Grade
This antibody recognizes histone H3 acetylated at lysines 9, 14, 18, 23 or 27 as confirmed by dot blot with non-modified histone H3 peptide or peptides No reaction with non-modified histone H3 peptide as tested by dot blot.
Tested applicationsSuitable for: ChIP, ICC/IF, WBmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse
Synthetic peptide corresponding to Human Histone H3 (N terminal) (acetyl K9 + K14 + K18 + K23 + K27).
- WB: HeLa cell lysate. ICC/IF: HeLa cells. ChIP: Chromatin prepared from HeLa cells.
Learn about ChIP assay kits, other ChIP antibodies, protocols and more in the ChIP assay guide.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.035% Sodium azide
Constituents: Whole serum, 30% Glycerol
ChIP Related Products
Our Abpromise guarantee covers the use of ab47915 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration. PubMed: 20080953
Use 10 µL per ChIP experiment
|WB||1/500 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).|
FunctionCore 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.
Sequence similaritiesBelongs to the histone H3 family.
Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
modificationsAcetylation 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.
Cellular localizationNucleus. Chromosome.
- Information by UniProt
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HP1α delocalises from chromocentres upon proteasome inhibition whereas the histone modifications remain unaffected.
(A) Proteasome inhibition does not affect canonical histone modifications on major and minor satellite repeats. NIH/3T3 (Mouse embryo fibroblast cell line) cells were treated with 20μM MG132 for 4h followed by ChIP–qPCR analysis using antibodies against repressive mark H3K9me3 and activating marks H3K4me3, H3K36me3, and H3ac (ab47915). Data is shown as relative enrichment to H3 with background subtraction. The y-axis scale was adjusted depending on the signal obtained with different antibodies. Error bars = SEM of 3 biological replicates.
ChIP was performed using a high sensitivity kit with 5 µg of chromatin from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and 10 µL of Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody ab47915.
ChIP DNA was used in qPCR with the negative control primer pairs or gene-specific primer pairs as indicated. Data are presented as binding events detected per 1000 cells using a normalization scheme which accounts for primer efficiency and the amount of chromatin used in the ChIP reaction.
All lanes : Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915) at 1/2000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, no treatment
Lane 2 : HeLa cells, treatment with sodium butyrate
Lysates/proteins at 10 µg per lane.
Predicted band size: 15 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?
ICC/IF image of ab47915 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum ab7481 / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47915, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, a goat anti-rabbit DyLight® 488 (IgG; H+L) used at a 1/250 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 1.43 µM.
ab47915 tested by dot blot analysis.
Dot blot analysis was used to confirm the specificity of ab47915. Acetylated peptides corresponding to the immunogen and related peptides were spotted onto PVDF and probed with the antibody at a dilution of 1:1,000. The amount of peptide (picomoles) spotted is indicated next to the row.
Lane 1: acetyl-Lys9 peptide. Lane 2: unmodified Lys9 peptide. Lane 3: acetyl-Lys14 peptide. Lane 4: unmodified Lys14 peptide. Lane 5: acetyl-Lys18 peptide. Lane 6: unmodified Lys18 peptide. Lane 7: acetyl-Lys23 peptide. Lane 8: unmodified Lys23 peptide. Lane 9: acetyl-Lys27 peptide. Lane 10: unmodified Lys27 peptide.
This product has been referenced in:
- Smith ER et al. TGF-ß1 modifies histone acetylation and acetyl-coenzyme A metabolism in renal myofibroblasts. Am J Physiol Renal Physiol N/A:N/A (2019). Read more (PubMed: 30623724) »
- Park HJ et al. HOS15 Interacts with the Histone Deacetylase HDA9 and the Evening Complex to Epigenetically Regulate the Floral Activator GIGANTEA. Plant Cell 31:37-51 (2019). Read more (PubMed: 30606777) »