Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915)
- Datasheet
- Specific References (8)
- Protocols
Overview
- 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
- SpecificityThis antibody recognizes histone H3 acetylated at lysines 9, 14, 18, 23 or 27 as confirmed by dot blot with nonmodified 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
- Immunogen
Synthetic peptide corresponding to Human Histone H3 (acetyl K9 + K14 + K18 + K23 + K27).
- Positive control
- WB: HeLa cell lysate. ICC/IF: HeLa cells.
Properties
- FormLiquid
- 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 - PurityWhole antiserum
- ClonalityPolyclonal
- IsotypeIgG
- Research areas
Associated products
- Compatible Secondaries
- Positive Controls
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Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| ChIP | Use at an assay dependent concentration. PubMed: 20080953 Use 10 µL per ChIP experiment |
|
| ICC/IF | 1/1000. | |
| WB | 1/500 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa). |
Target
- 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.
- Post-translational
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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Database links
- Entrez Gene: 8290 Human
- Entrez Gene: 8350 Human
- Entrez Gene: 8351 Human
- Entrez Gene: 8352 Human
- Entrez Gene: 8353 Human
- Entrez Gene: 8354 Human
- Entrez Gene: 8355 Human
- Entrez Gene: 8356 Human
see all -
Alternative names
- H3.3A antibody
- HIST1 cluster, H3E antibody
- H3 histone family, member A antibody
see all
Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade images
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ChIP - Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915)Histone H3ac (pan-acetyl) antibody tested by ChIP. ChIP was performed using a High Sensitivity Kit with 5 µg of chromatin from HeLa cells and 10 µl of Histone H3ac antibody. 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.
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Western blot - Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody (ab47915)All lanes : Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915) at 1/2000 dilution
Lane 1 : HeLa 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?) -
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915)ICC/IF image of ab47915 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 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 (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 1.43µM.
References for Anti-Histone H3 (acetyl K9 + K14 + K18 + K23 + K27) antibody - ChIP Grade (ab47915)
This product has been referenced in:
- Su LC et al. Isolation and characterization of an osmotic stress and ABA induced histone deacetylase in Arachis hygogaea. Front Plant Sci 6:512 (2015). Read more (PubMed: 26217363) »
- Kim KB et al. H3K9 methyltransferase G9a negatively regulates UHRF1 transcription during leukemia cell differentiation. Nucleic Acids Res 43:3509-23 (2015). Human . Read more (PubMed: 25765655) »
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