There was no cross-reactivity observed with recombinant H3 or the following modifications Acetyl-K9/pS10, -K18, -K23, and -K27 in dot plot.
We have data to indicate that this antibody will not cross react with mouse. The antibody was tested against brain, kidney, spleen and heart lysates.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
IHC - Wholemount
Use at an assay dependent concentration.
Use at an assay dependent concentration. PubMed: 19188451
1/2000. Detects a band of approximately 11 kDa (predicted molecular weight: 11 kDa).
1/100 - 1/250.
1/250 - 1/500.
Is unsuitable for Flow Cyt.
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.
Belongs to the histone H3 family.
Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
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.
Immunocytochemistry/Immunofluorescence analysis of untreated HeLa cells and TSA (500ng/ml, 4h) and treated HeLa cells labelling Histone H3 (acetyl K14) with purified ab52946 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100, counterstained with ab150120 AlexaFluor®594 Goat anti-Mouse secondary 1:1000 (2ug/ml). An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Negative Control 1: Rabbit primary antibody and anti-mouse secondary antibody(ab150120)
Negative Control 2: Mouse primary antibody(ab7291) and anti-rabbit secondary antibody(ab150077)
Western blot - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
All lanes : Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946) at 1/2000 dilution
Lane 1 : C6 cell lysates untreated Lane 2 : C6 cell lysates treated with TSA
Lysates/proteins at 10 µg per lane.
Secondary All lanes : goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 11 kDa Observed band size: 11 kDa
ab52946 at 1/100-1/250 dilution staining Histone H3 (acetyl K14) in human adenocarcinoma of uterus by Immunohistochemistry, Paraffin embedded tissue.
Immunoprecipitation - Anti-Histone H3 (acetyl K14) [EP964Y] antibody - ChIP Grade (ab52946)This image is courtesy of an anonymous Abreview
ab52946 immunoprecipitating Histone H3 (acetyl K14) in Drosophila melanogaster embryo whole tissue lysate. 500µg of tissue lysate was incubated with the undiluted primary antibody and matrix (Protein G) for 2 hours at 4°C. For western blotting ab52946 (1/20000) was used to confirm successful immunoprecipation.
Anderson L et al. Histone deacetylase inhibition modulates histone acetylation at gene promoter regions and affects genome-wide gene transcription in Schistosoma mansoni. PLoS Negl Trop Dis11:e0005539 (2017).
Read more (PubMed: 28406899) »
Zhang T et al. UHRF2 decreases H3K9ac expression by interacting with it through the PHD and SRA/YDG domain in HepG2 hepatocellular carcinoma cells. Int J Mol Med39:126-134 (2017).
Read more (PubMed: 28004105) »