Anti-Histone H3 (acetyl K56) [EPR996Y] antibody (ab76307)
- Product nameAnti-Histone H3 (acetyl K56) [EPR996Y] antibodySee all Histone H3 primary antibodies ...
- DescriptionRabbit monoclonal [EPR996Y] to Histone H3 (acetyl K56)
- Tested applicationsWB, IHC-P, ICC/IF more details
- Species reactivityReacts with: Mouse, Rat, Human, Saccharomyces cerevisiae, African Green Monkey
A synthetic acetylated peptide corresponding to residues surrounding Lys 56 of Histone H3
- Positive control
- C6 cell lysate, untreated or treated with TSA; HeLa cells.
- General notes
Produced under U.S. Patent No. 5,675,063.
This product is available conjugated to Alexa Fluor® 488 see ab170098.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
- PurityTissue culture supernatant
- Clonality Monoclonal
- Clone numberEPR996Y
- Research Areas
Our Abpromise guarantee covers the use of ab76307 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Use of HRP-conjugated or polymerized HRP secondary antibodies, stronger signals have been found using the polymerized HRP secondary.
WB: 1/1000 - 1/2000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
Is unsuitable for Flow Cyt or IP.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
- 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.
- 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
- Entrez Gene: 100364501 Rat
- Entrez Gene: 100365669 Rat
- Entrez Gene: 291159 Rat
- Entrez Gene: 679994 Rat
- Entrez Gene: 680511 Rat
- Entrez Gene: 682330 Rat
- Omim: 142780 Human
- Omim: 601058 Human
- Omim: 601128 Human
- SwissProt: P68431 Human
- SwissProt: P84243 Human
- SwissProt: Q71DI3 Human
- SwissProt: P68433 Mouse
- SwissProt: Q6LED0 Rat
- 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
- Unigene: 138090 Rat
- H 3 antibody
- H3 antibody
- H3/A antibody
- H31_HUMAN antibody
- H3FA antibody
- HIST1H3J antibody
- Histone cluster 1, H3a antibody
- Histone H3.1 antibody
- Histone H3.3 antibody
- Histone H3/a antibody
- Histone H3/b antibody
- Histone H3/c antibody
- Histone H3/d antibody
- Histone H3/f antibody
- Histone H3/h antibody
- Histone H3/i antibody
- Histone H3/j antibody
- Histone H3/k antibody
- Histone H3/l antibody
Anti-Histone H3 (acetyl K56) [EPR996Y] antibody images
ab76307 staining Histone H3 (HEK56 Ac) in African Green Monkey kidney epithelial (CV-1 line) cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton and blocked with 5% BSA for 1 hour at 21°C. Samples were incubated with primary antibody (1/500) for 16 hour at 4°C.
CV-1 cells were infected with Simian virus 40 (SV40). 32 hours later the cells were incubated with EdU for 30 minutes before pre-extraction to detect active DNA replication foci and fixed, stained, and visualized by confocal microscopy.
Blue - Hoechst stain, red - anti-T-Ag, magenta - EdU, green - anti-H3K56 Ac. Bar, 5 µm.
ab76307 at 1/100 dilution staining Histone H3 in HeLa cells by Immunofluorescence.
All lanes : Anti-Histone H3 (acetyl K56) [EPR996Y] antibody (ab76307) at 1/2000 dilution
Lane 1 : C6 cell lysate, untreated
Lane 2 : C6 cell lysate, treated with TSA
Lysates/proteins at 10 µg per lane.
goat anti-rabbit HRP at 1/1000 dilution
Predicted band size : 15 kDa
Observed band size : 15 kDa
IHC image of ab76307 staining in Breast Cancer 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 ab76307, undiluted, 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.
ab76307 showing positive staining in Cervical carcinoma tissue.
ab76307 showing positive staining in Gastric adenocarcinoma tissue.
ab76307 showing positive staining in Lung adenocarcinoma tissue.
ab76307 showing positive staining in Lymphoma tissue.
References for Anti-Histone H3 (acetyl K56) [EPR996Y] antibody (ab76307)
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). WB ; Human . Read more (PubMed: 23415232) »
- Kato H et al. Spt6 prevents transcription-coupled loss of posttranslationally modified histone H3. Sci Rep 3:2186 (2013). Read more (PubMed: 23851719) »