Anti-Histone H3 (phospho T6) antibody - ChIP Grade (ab14102)

Overview

  • Product nameAnti-Histone H3 (phospho T6) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
  • Description
    Rabbit polyclonal to Histone H3 (phospho T6) - ChIP Grade
  • Tested applicationsChIP, IHC-P, WB, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, phosphorylated at T6.

  • Positive control
    • Colcemid-treated HeLa histone Preparation; Hela whole cell extract

Properties

Applications

Our Abpromise guarantee covers the use of ab14102 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
ChIP
IHC-P
WB
ICC/IF
  • Application notesChIP: Use at an assay dependent dilution (PMID 20228790).
    ICC/IF: 1/200.
    IHC-P: Use at an assay dependent dilution.
    WB: 1/10000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • FunctionVariant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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 throughout the cell cycle independently of DNA synthesis.
    • Post-translational
      modifications
      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.
      Specifically enriched in modifications associated with active chromatin such as methylation at Lys-5 (H3K4me), Lys-37 and Lys-80. 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), which are linked to gene repression, are underrepresented. 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. Phosphorylation on Ser-32 (H3S31ph) is specific to regions bordering centromeres in metaphase chromosomes.
      Ubiquitinated. Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination.
    • Cellular localizationNucleus. Chromosome.
    • Target information above from: UniProt accession P84243 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links
    • Alternative names
      • H3 3 like sequence MH921 antibody
      • H3 3 like sequence MH921 antibody
      • H3 3A antibody
      • H3 a antibody
      • H3 b antibody
      • H3 c antibody
      • H3 d antibody
      • H3 f antibody
      • H3 h antibody
      • H3 histone family member E pseudogene antibody
      • H3 histone family member E pseudogene antibody
      • H3 i antibody
      • H3 j antibody
      • H3 k antibody
      • H3 l antibody
      • H33_HUMAN antibody
      • H3F3 antibody
      • H3f3b antibody
      • Histone H3 3 pseudogene antibody
      • Histone H3 3 pseudogene antibody
      • Histone H3.3 antibody
      see all

    Anti-Histone H3 (phospho T6) antibody - ChIP Grade images

    • All lanes : Anti-Histone H3 (phospho T6) antibody - ChIP Grade (ab14102) at 1 µg/ml

      Lane 1 : Colcemid treated histone
      Lane 2 : Untreated histone
      Lane 3 : Colcemid treated histone with Histone H3 peptide- phospho T6 (ab14103) at 1 µg/ml
      Lane 4 : Untreated histone with Histone H3 peptide- phospho T6 (ab14103) at 1 µg/ml
      Lane 5 : Colcemid treated histone with Histone H3 peptide - unmodified (ab2903) at 1 µg/ml
      Lane 6 : Untreated histone with Histone H3 peptide - unmodified (ab2903) at 1 µg/ml
      Lane 7 : Colcemid treated histone with Histone H3 peptide - phospho T3 (ab17578)
      Lane 8 : Untreated histone with Histone H3 peptide - phospho T3 (ab17578)
      Lane 9 : Colcemid treated histone with Histone H3 peptide - phospho T11 (ab24444)
      Lane 10 : Untreated histone with Histone H3 peptide - phospho T11 (ab24444)

      Lysates/proteins at 2.5 µg per lane.


      Predicted band size : 15 kDa
      Observed band size : 17 kDa (why is the actual band size different from the predicted?)
    • ab14102 staining Histone H3 in HeLa cells, by ICC/IF, in conjunction with a secondary goat anti-rabbit antibody conjugated to Cy3® (green). Cells counterstained with DAPI to show the nucleus (red).

      See Abreview

    References for Anti-Histone H3 (phospho T6) antibody - ChIP Grade (ab14102)

    This product has been referenced in:
    • Metzger E  et al. Phosphorylation of histone H3T6 by PKCbeta(I) controls demethylation at histone H3K4. Nature 464:792-6 (2010). WB, IHC-P, ChIP ; Human . Read more (PubMed: 20228790) »
    • Hurd PJ  et al. Phosphorylation of histone H3 Thr-45 is linked to apoptosis. J Biol Chem 284:16575-83 (2009). WB ; Human . Read more (PubMed: 19363025) »

    See all 3 Publications for this product

    Product Wall

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (pancreatic cancer)
    Specification pancreatic cancer
    Fixative Paraformaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
    Permeabilization Yes - PBST
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 11 2011

    Application Immunocytochemistry/ Immunofluorescence
    Sample Mouse Cell (Mouse pancreatic cancer primary cell lines)
    Specification Mouse pancreatic cancer primary cell lines
    Fixative Acetone:Methanol 1:1
    Permeabilization Yes - PBST
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 01 2010

    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Paraformaldehyde
    Username

    Dr. Kirk McManus

    Verified customer

    Submitted Nov 02 2006

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"