Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441)
Overview
- Product nameAnti-Histone H3 (acetyl K9) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
- DescriptionRabbit polyclonal to Histone H3 (acetyl K9) - ChIP Grade
- SpecificityIn Dot blot detects 50ng of mono-acetylated peptide corresponding to position Lys9 in the N-terminal sequence of Histone H3. Does not detect the mono-acetylated peptide corresponding to acetyl-lysine at position 14 or unacetylated Histone H3.
- Tested applicationsICC/IF, CHIPseq, Dot Blot, WB, ChIP more details
- Species reactivityReacts with: Mouse, Human, Tetrahymena sp., Xenopus laevis, Caenorhabditis elegans, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, Toxoplasma gondii
Predicted to work with: Saccharomyces cerevisiae - Immunogen
Synthetic peptide: ARTKQTARAcKSTG-C conjugated to KLH, corresponding to amino acids 1-12 of Human Histone H3.
- Positive controlWB: TSA-treated and acid extracted HeLa and NIH/3T3 cells. ICC:polytene chromosomes of Drosophila melanogaster
Properties
- FormLiquid
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage buffer0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide - made up to 30% glycerol
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Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab4441 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | ICC/IF: 1/100 - 1/200. |
| CHIPseq | CHIPseq: Use at an assay dependent dilution. |
| Dot Blot | Dot: Use a concentration of 0.5 µg/ml. Detects 50ng of mono-acetylated peptide corresponding to position Lys9 in the N-terminal sequence of Histone H3. Does not detect the mono-acetylated peptide corresponding to acetyl-lysine at position 14 or unacetylated Histone H3. |
| WB | WB: 1/500. Detects a band of approximately 17 kDa. |
| ChIP | ChIP: Use 2µg for 106 cells. |
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.
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Database links
- 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
- Entrez Gene: 8357 Human
- Entrez Gene: 8358 Human
- Entrez Gene: 8968 Human
- Entrez Gene: 319152 Mouse
- Entrez Gene: 319153 Mouse
- Entrez Gene: 360198 Mouse
- Entrez Gene: 97908 Mouse
- Omim: 601128 Human
- Omim: 142780 Human
- Omim: 601058 Human
- SwissProt: P84243 Human
- SwissProt: Q71DI3 Human
- SwissProt: P68431 Human
- SwissProt: P68433 Mouse
- Unigene: 132854 Human
- Unigene: 247813 Human
- Unigene: 247814 Human
- Unigene: 248176 Human
- Unigene: 443021 Human
- Unigene: 484990 Human
- Unigene: 532144 Human
- Unigene: 533292 Human
- Unigene: 546315 Human
- Unigene: 586261 Human
- Unigene: 591778 Human
- Unigene: 221301 Mouse
- Unigene: 261657 Mouse
- Unigene: 377874 Mouse
- Unigene: 390558 Mouse
- Unigene: 397328 Mouse
see all
Target information above from: UniProt accession
P68431
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- H 3 antibodyH3 3 Like Sequence MH921 antibodyH3 3A antibody
- H3 3A antibodyH3 antibodyH3 Histone antibodyH3 Histone Family Member E Pseudogene antibodyH3.4 antibodyH3/A antibodyH3/g antibodyH31_HUMAN antibodyH3F3 antibodyH3F3 antibodyH3FA antibodyH3FT antibodyH3t antibodyHIST1H3J antibodyHIST3H3 antibodyHIST3H3 antibodyHistone cluster 1, H3a antibodyHistone H3 3 Pseudogene antibodyHistone H3.1 antibodyHistone H3.3 antibodyHistone H3/a antibodyHistone H3/b antibodyHistone H3/c antibodyHistone H3/d antibodyHistone H3/f antibodyHistone H3/h antibodyHistone H3/i antibodyHistone H3/j antibodyHistone H3/k antibodyHistone H3/l antibody
see all
Anti-Histone H3 (acetyl K9) antibody - ChIP Grade images
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ChIP - Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441)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 ab4441 (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. -
Immunocytochemistry/ Immunofluorescence - Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441)This image was submitted as part of a review by review by Anita Ciurciu.Fruit fly (Drosophila melanogaster) Cell (polytene chromosomes) were fixed in formaldehyde, blocked in 1% BSA for 30 minutes and incubated with ab4441 (1/100) for 12 hours.
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Western blot - Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441)All lanes : Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441) at 0.5 µg/ml
Lane 1 : Untreated HeLa cell acid-extract
Lane 2 : HeLa cell acid-extract treated with sodium butyrate
References for Anti-Histone H3 (acetyl K9) antibody - ChIP Grade (ab4441)
This product has been referenced in:
- Tropberger P et al. Regulation of transcription through acetylation of H3K122 on the lateral surface of the histone octamer. Cell 152:859-72 (2013). ChIP ; Human . Read more (PubMed: 23415232) »
- Rajabi H et al. MUC1-C oncoprotein induces TCF7L2 transcription factor activation and promotes cyclin D1 expression in human breast cancer cells. J Biol Chem 287:10703-13 (2012). ChIP ; Human . Read more (PubMed: 22318732) »
