Anti-Histone H3 (acetyl K122) antibody - ChIP Grade (ab33309)

Overview

  • Product nameAnti-Histone H3 (acetyl K122) antibody - ChIP GradeSee all Histone H3 primary antibodies ...
  • Description
    Rabbit polyclonal to Histone H3 (acetyl K122) - ChIP Grade
  • Tested applicationsIP, ChIP, CHIPseq, WB, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Human, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster), Brassica oleracea
    Predicted to work with: a wide range of other species
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human Histone H3, acetylated at K122.

    (Peptide available as ab34466.)

  • Positive control
    • ab33309 gave a positive result in HeLa Histone Preparation Nuclear Lysate - Butyrated treated.

Properties

Applications

Our Abpromise guarantee covers the use of ab33309 in the following tested applications.

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

Application Notes
IP Use at an assay dependent concentration. PubMed: 23415232
ChIP Use at an assay dependent concentration. PubMed: 23415232
CHIPseq Use at an assay dependent concentration. PubMed: 23415232
WB Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 (acetyl K122) peptide (ab34466).
ICC/IF Use a concentration of 1 µg/ml.

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
    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.
    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
  • Alternative names
    • H 3 antibody
    • H3 3 Like Sequence MH921 antibody
    • H3 3A antibody
    • H3 antibody
    • H3 Histone antibody
    • H3 Histone Family Member E Pseudogene antibody
    • H3.4 antibody
    • H3/A antibody
    • H3/g antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3FA antibody
    • H3FT antibody
    • H3t antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • Histone cluster 1, H3a antibody
    • Histone H3 3 Pseudogene 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
    see all

Anti-Histone H3 (acetyl K122) antibody - ChIP Grade images

  • All lanes : Anti-Histone H3 (acetyl K122) antibody - ChIP Grade (ab33309) at 1 µg/ml

    Lane 1 : HeLa Histone Preparation Nuclear Lysate - Butyrated
    Lane 2 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Human Histone H3 (acetyl K122) peptide (ab34466) at 1 µg/ml
    Lane 3 : HeLa Histone Preparation Nuclear Lysate - Butyrated with Histone H3 peptide - unmodified (ab34467) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 11.2 kDa (possible cross reactivity).
  • ICC/IF image of ab33309 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33309, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

References for Anti-Histone H3 (acetyl K122) antibody - ChIP Grade (ab33309)

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, IP, ChIP, ICC/IF, CHIPseq ; Mouse, Human, Fruit fly (Drosophila melanogaster) . Read more (PubMed: 23415232) »

See 1 Publication for this product

Product Wall

I am sorry this antibody has not been working for you in a guaranteed species and application. If we cannot get this antibody to work for you I would be happy to either refund or replace the product free of charge. After reviewing your protocol I have ...

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As I mentioned in my previous email, I have looked into the quality control data for the three lots of antibody you have been using. They all passed our testing and from the results (summarised in the attachment of this email), all seem to be equally e...

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Thank you for contacting us.

Unfortunately Kate is away but I will try to deal with your case as best I can.
Currently we only have stock of lot number GR38318-1, the same as you have already tried. I am concerned at the variable result...

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Application Immunocytochemistry/ Immunofluorescence
Sample Arabidopsis thaliana Cell (Root tip interphase nuclei)
Specification Root tip interphase nuclei
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Feb 02 2012

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (15)
Sample Brassica oleracea Cell lysate - nuclear (inflorescences)
Specification inflorescences
Treatment SODIUM BUTYRATE 10mM
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Mr. walid mahrez

Verified customer

Submitted Jan 21 2012

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