Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)

Overview

• Product nameAnti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP GradeSee all Histone H3 primary antibodies ...
• Description
  Rabbit monoclonal [EP964Y] to Histone H3 (acetyl K14) - ChIP Grade
• SpecificityThere 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.
• Tested applicationsChIP, WB, IP, IHC-P, ICC/IF more details
• Species reactivity
  Reacts with: Rat, Human, Saccharomyces cerevisiae, Schizosaccharomyces pombe
  

  Does not react with

  Mouse
• Immunogen

  A synthetic acetyl-specific peptide corresponding to residues around Lysine 14 of Histone H3

• Positive controlC6 cell lysates, human adenocarcinoma of uterus tissue and HeLa cells.
• General notesProduced under U.S. Patent No. 5,675,063.

Properties

• FormLiquid
• Storage instructionsStore at -20°C. Stable for 12 months at -20°C
• Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
• PurityTissue culture supernatant
• Clonality Monoclonal
• Clone numberEP964Y
• IsotypeIgG
• Research Areas
  • Epigenetics and Nuclear Signaling
  • ChIP'ing antibodies
  • ChIP'ing antibodies

  • Epigenetics and Nuclear Signaling
  • Histones
  • H3
  • Acetylated

Applications

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.

Target information above from: UniProt accession P68431 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• 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: 100364501 Rat
  • Entrez Gene: 100365669 Rat
  • Entrez Gene: 291159 Rat
  • Entrez Gene: 679994 Rat
  • Entrez Gene: 680511 Rat
  • Entrez Gene: 682330 Rat
  • Omim: 601128 Human
  • Omim: 142780 Human
  • Omim: 601058 Human
  • SwissProt: P84243 Human
  • SwissProt: Q71DI3 Human
  • SwissProt: P68431 Human
  • 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: 138090 Rat

  see all

• Alternative names
  H 3 antibodyH3 antibodyH3/A antibody

  H31_HUMAN antibodyH3FA antibodyH4F antibodyHIST1H3J antibodyHIST1H4 antibodyHistone cluster 1, H3a 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 antibodyHistone H4 antibody

  see all

Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade images

• 

  Western blot - Histone H3 (acetyl K14) antibody [EP964Y] (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
  goat anti-rabbit HRP at 1/2000 dilution
  
  Predicted band size : 11 kDa
  Observed band size : 11 kDa
  
• 

  Immunohistochemistry (Paraffin-embedded sections) - Histone H3 (acetyl K14) antibody [EP964Y] (ab52946)
  ab52946 at 1/100-1/250 dilution staining Histone H3 (acetyl K14) in human adenocarcinoma of uterus by Immunohistochemistry, Paraffin embedded tissue.
• 

  Immunocytochemistry/ Immunofluorescence - Histone H3 (acetyl K14) antibody [EP964Y] (ab52946)
  ab52946 at 1/250-1/500 dilution staining Histone H3 (acetyl K14) in HeLa cells by Immunofluorescence.
• 

  Immunocytochemistry/ Immunofluorescence - Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (ab52946)
  ICC/IF image of ab52946 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 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 (ab52946, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) 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.